Compounds for treating proliferative disorders

ABSTRACT

Disclosed are compounds of Formula (I) and methods of using compounds of the invention for treating a subject with a proliferative disorder, such as cancer, and methods for treating disorders responsive to Hsp70 induction and/or natural killer induction. Also, disclosed are pharmaceutical compositions comprising compounds of the invention and a pharmaceutically acceptable carrier.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.61/066,526, filed on Feb. 21, 2008. The entire teachings of the aboveapplication is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Heat shock proteins (HSPs) are found in virtually all prokaryotic andeukaryotic cells where they support folding of nascent polypeptides,prevent protein aggregation, and assist transport of other proteinsacross membranes. The proteins in the Hsp70 family (referred tocollectively as “Hsp70”) play a dual role of protecting cells fromlethal damage after environmental stress, on the one hand, and targetingcells for immune mediated cytolytic attack on the other hand. Increasedexpression of Hsp70 in the cytoplasma is known to protect a broad rangeof cells under stress by preventing the misfolding, aggregation anddenaturation of cytoplasmic proteins and inhibiting various apoptoticpathways (Mosser, et al., Mol Cell Biol. 2000 October; 20(19):7146-7159; Yenari, Adv Exp Med Biol, 2002, 513, 281-299; Kiang andTsokos, Pharmacol Ther. 1998; 80(2):182-201). However, membrane-boundHsp70 provides a target structure for cytolytic attack mediated bynatural killer cells.

Cells can experience stress due to temperature; injury (trauma); geneticdisease; metabolic defects; apoptosis; infection; toxins; radiation;oxidants; excess/lack of nutrients or metabolic products; and the like.For example, it is known in the art that cells damaged in the followingvariety of medical conditions can experience a protective effect inresponse to Hsp70.

Protein misfolding/aggregation conditions resulting in neurodegenerationinclude Alzheimer's disease (Zhang, et al., J. Neuroscience, 2004,24(23), 5315-5321; Klettner, Drug News Perspect, 2004 17(5), 299-306);Huntington's disease (Klettner, ibid); Parkinson's disease (Auluck, etal., Science, 2002, 295(5556), 865-868); and the like. Otherneurodegenerative conditions include spinal/bulbar muscular atrophy(Sobue, Nihon Shinkei Seishin Yakurigaku Zasshi, 2001, 21(1), 21-25);and familial amyotrophic lateral sclerosis (Howland, et al., Proc NatAcad Sci USA, 2002, 99(3), 1604-1609; Sobue, ibid; Vleminck, et al., JNeuropathol Exp Neurol, 2002, 61(11), 968-974).

Ischemia and associated oxidative damage affects diverse tissuesincluding: neurons and glia (Carmel, et al., Exp Neurol, 2004, 185(1)81-96; Renshaw and Warburton, Front Biosci, 2004, 9, 110-116; Yenari,Adv Exp Med Biol, 2002, 513, 281-299; Kelly and Yenari, Curr Res MedOpin, 2002, 18 Suppl 2, s55-60; Lee, et al., Exp Neurol, 2001, 170(1),129-139; Klettner, ibid; Klettner and Herdegen, Br J Pharmacol, 2003,138(5), 1004-1012); cardiac muscle (Marber, M. S., et al. (1995) J.Clin. Invest. 95:1446-1456; Plumier, J. C., et al. (1995) J. Clin.Invest. 95:1854-1860; Radford, N. B., et al. (1996) Proc. Natl. Acad.Sci. USA 93(6): 2339-2342; Voss, et al., Am J Physiol Heart Circ Physiol285: H687-H692, 2003); liver tissue (Doi, et al.,Hepatogastroenterology. 2001 March-April; 48(38):533-40; Gao, et al.World J Gastroenterol 2004; 10(7):1019-1027); skeletal muscle (Lepore etal., Cell Stress & Chaperones, 2001, 6(2), 93-96); kidney tissue (Chen,et al., Kidney Int. 1999; 56: 1270-1273; Beck, et al., Am J PhysiolRenal Physiol 279: F203-F215, 2000.); pulmonary tissue (Hiratsuka, etal., J Heart Lung Transplant. 1998 December; 17(12):1238-46); pancreatictissue (Bellmann, et al., J Clin Invest. 1995 June; 95(6): 2840-2845),and the like.

Seizure conditions that damage neurons include, e.g., epileptic seizure(Yenari, ibid; Blondeau, et al. Neuroscience 2002, 109(2), 231-241); orchemically induced seizure (Tsuchiya, et al., Neurosurgery, 2003, 53(5),1179-1187).

Thermal stresses include hyperthermia conditions such as fever, heatstroke, and the like (Barclay and Robertson, J Neurobiol, 2003 56(4),360-271; Sato, et al., Brain Res, 1996, 740(1-2), 117-123); andhypothermia (Kandor and Goldberg, Proc Natl Acad Sci U S A. 1997 May 13;94(10): 4978-4981).

Aging includes conditions such as atherosclerosis which affects smoothmuscle cells (Minowada, G. and Welch, W. J. (1995) J. Clin. Invest.95:3-12; Johnson, A. J., et al. (1995) Arterio. Thromb. Vasc. Biol.15(1):27-36).

Other conditions include radiation damage, e.g., from ultraviolet lightto tissues such as murine fibroblasts (Simon, M. M., et al. (1995) J.Clin. Res. 95(3): 926-933), and light damage to retinal cells (Yu, et,al, Molecular Vision 2001; 7:48-56).

Trauma includes, for example, mechanical injury, e.g., pressure damageto retinal ganglions in glaucoma (Ishii, et al., Invest Opthalmol V isSci, 2003, 44(5), 1982-1992).

Toxic conditions include doses of chemicals or biochemicals, forexample, methamphetamine (Malberg & Seiden, Poster “MDMA AdministrationInduces Expression of HSP70 in the Rat Brain” Society for NeuroscienceAnnual Meeting, New Orleans, La., Oct. 25-30, 1997); antiretroviral HIVtherapeutics (Keswani, et al., Annals Neurology, 2002, 53(1), 57-64);heavy metals, amino acid analogs, chemical oxidants, ethanol, glutamate,and other toxins (Ashburner, M. and Bonner, J. J. (1979) Cell:17:241-254; Lindquist, S. (1986) Ann. Rev. Biochem. 55:1151-1191; Craig,E. A. (1985) Crit. Rev. Biochem. 18(3):239-280; Morimoto, et al., In:The Biology of Heat Shock Proteins and Molecular Chaperone, (1994) pp.417-455. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, N.Y.);and the like.

Cystic fibrosis is a genetic disorder which results from a mutation in asingle glycoprotein called the cystic fibrosis transmembrane conductanceregulator (CFTR). As a result of the mutation, post-translationalprocessing of CFTR cannot proceed correctly and the glycoprotein failsto be delivered to the cell membrane. Induction of Hsp70 has been shownto overcome this defective processing and results in functional CFTRprotein on the cell surface (Choo-Kang and Zeitlin, Am. J. Physiol. LungCell Mol. Physiol. (2001), 281:L58-L68).

Therefore, there is a need for new methods of increasing expression ofHsp70 in order to treat disorders responsive to Hsp70.

Extracellular Hsp70 and membrane bound Hsp70 have been shown to play keyroles in activation of the innate immune system. Monocytes have beenshown to secrete proinflammatory cytokines in response to soluble Hsp70protein and membrane bound Hsp70 has been shown to provide a targetstructure for cytolytic attack by natural killer cell.

Natural killer (NK) cells, a type of white blood cell, are known to bean important component of the body's immune system. Because the definingfunction of NK cells is spontaneous cytotoxicity without priorimmunization, NK cells can be the first line of defense in the immunesystem, and are believed to play a role in attacking cancer cells andinfectious diseases. Many conditions, such as immunodeficiency diseases,aging, toxin exposure, endometriosis, and the like can leave subjectswith lowered NK cell activity or dysfunctional NK cells.

For example, subjects can have decreased or deficient NK cell activity,in conditions such as chronic fatigue syndrome (chronic fatigue immunedysfunction syndrome) or Epstein-Barr virus, post viral fatiguesyndrome, post-transplantation syndrome or host-graft disease, exposureto drugs such as anticancer agents or nitric oxide synthase inhibitors,natural aging, and various immunodeficiency conditions such as severecombined immunodeficiency, variable immunodeficiency syndrome, and thelike. (Caligiuri M, Murray C, Buchwald D, Levine H, Cheney P, PetersonD, Komaroff A L, Ritz J. Phenotypic and functional deficiency of naturalkiller cells in patients with chronic fatigue syndrome. Journal ofImmunology 1987; 139: 3306-13; Morrison L J A, Behan W H M, Behan P O.Changes in natural killer cell phenotype in patients with post-viralfatigue syndrome. Clinical and Experimental Immunology 1991; 83: 441-6;Klingemann, HG Relevance and Potential of Natural Killer Cells in StemCell Transplantation Biology of Blood and Marrow Transplantation 2000;6:90-99; Ruggeri L, Capanni M, Mancusi A, Aversa F, Martelli M F,Velardi A. Natural killer cells as a therapeutic tool in mismatchedtransplantation. Best Pract Res Clin Haematol. 2004 September;17(3):427-38; Cifone M G, Ulisse S, Santoni A. Natural killer cells andnitric oxide. Int Immunopharmacol. 2001 August; 1(8):1513-24; Plackett TP, Boehmer E D, Faunce D E, Kovacs E J. Aging and innate immune cells. JLeukoc Biol. 2004 August; 76(2):291-9. Epub 2004 Mar. 23; Alpdogan O,van den Brink M R. IL-7 and IL-15: therapeutic cytokines forimmunodeficiency. Trends Immunol. 2005 January; 26(1):56-64; Heusel J W,Ballas Z K. Natural killer cells: emerging concepts in immunity toinfection and implications for assessment of immunodeficiency. Curr OpinPediatr. 2003 December; 15(6):586-93; Hacein-Bey-Abina S, Fischer A,Cavazzana-Calvo M. Gene therapy of X-linked severe combinedimmunodeficiency. Int J Hematol: 2002 November; 76(4):295-8; Baumert E,Schlesier M, Wolff-Vorbeck G, Peter H H. Alterations in lymphocytesubsets in variable immunodeficiency syndrome Immun Infekt. 1992 July;20(3):73-5.)

NK cells are known to have activity against a wide range of infectiouspathogens such as bacteria, viruses, fungi, protozoan parasites,combined infections, e.g., combined bacterial/viral infections, and thelike. NK cells are believed to be particularly important in combatingintracellular infections where the pathogens replicate in the subjectscells, e.g., a substantial fraction of viruses and many other pathogensthat can form intracellular infections.

For example, a wide range of fungal infections are reported to betargeted by NK cells such as Cryptococcus neoformans, dermatophytes,e.g., Trichophyton rubrum, Candida albicans, Coccidioides immitis,Paracoccidioides brasiliensis, or the like (Hidore M R, Mislan T W,Murphy J W. Responses of murine natural killer cells to binding of thefungal target Cryptococcus neoformans Infect Immun. 1991 April;59(4):1489-99; Akiba H, Motoki Y, Satoh M, Iwatsuki K, Kaneko F;Recalcitrant trichophytic granuloma associated with NK-cell deficiencyin a SLE patient treated with corticosteroid. Eur J. Dermatol. 2001January-February; 11(1):58-62; Mathews H L, Witek-Janusek L. Antifungalactivity of interleukin-2-activated natural killer (NK1.1+) lymphocytesagainst Candida albicans. J Med. Microbiol. 1998 November;47(11):1007-14; Ampel N M, Bejarano G C, Galgiani J N. Killing ofCoccidioides immitis by human peripheral blood mononuclear cells. InfectImmun. 1992 October; 60(10):4200-4; Jimenez B E, Murphy J W. In vitroeffects of natural killer cells against Paracoccidioides brasiliensisyeast phase. Infect Immun. 1984 November; 46(2):552-8.)

Also targeted by NK cells are bacteria, especially intracellularbacteria, e.g., Mycobacterium tuberculosis, Mycobacterium avium,Listeria monocytogenes, many different viruses, such as humanimmunodeficiency virus, herpesviruses, hepatitis, and the like, andviral/bacterial co-infection (Esin S, Batoni G, Kallenius G, Gaines H,Campa M, Svenson S B, Andersson R, Wigzell H. Proliferation of distincthuman T cell subsets in response to live, killed or soluble extracts ofMycobacterium tuberculosis and Myco. avium. Clin Exp Immunol. 1996 June;104(3):419-25; Kaufmann S H. Immunity to intracellular bacteria. AnnuRev Immunol. 1993; 11:129-63; See D M, Khemka P, Sahl L, Bui T, Tilles JG. The role of natural killer cells in viral infections. Scand J.Immunol. 1997 September; 46(3):217-24; Brenner B G, Dascal A, MargoleseR G, Wainberg M A. Natural killer cell function in patients withacquired immunodeficiency syndrome and related diseases. J Leukoc Biol.1989 July; 46(1):75-83; Kottilil S, Natural killer cells in HIV-1infection: role of NK cell-mediated non-cytolytic mechanisms inpathogenesis of HIV-1 infection. Indian Exp Biol. 2003 November;41(11):1219-25; Herman R B, Koziel M J. Natural killer cells andhepatitis C: is losing inhibition the key to clearance? ClinGastroenterol Hepatol. 2004 December; 2(12):1061-3; Beadling C, Slifka MK. How do viral infections predispose patients to bacterial infections?Curr Opin Infect Dis. 2004 June; 17(3):185-91)

In addition, NK cells combat protozoal infections includingtoxoplasmosis, trypanosomiasis, leishmaniasis and malaria, especiallyintracellular infections (Korbel D S, Finney O C, Riley E M. Naturalkiller cells and innate immunity to protozoan pathogens. Int JParasitol. 2004 December; 34(13-14):1517-28; Ahmed J S, Mehlhorn H.Review: the cellular basis of the immunity to and immunopathogenesis oftropical theileriosis. Parasitol Res. 1999 July; 85(7):539-49; Osman M,Lausten S B, El-Sefi T, Boghdadi I, Rashed M Y, Jensen S L. Biliaryparasites. Dig Surg. 1998; 15(4):287-96; Gazzinelli R T, Denkers E Y,Sher A. Host resistance to Toxoplasma gondii: model for studying theselective induction of cell-mediated immunity by intracellularparasites. Infect Agents Dis. 1993 June; 2(3):139-49; Askonas B A,Bancroft G J. Interaction of African trypanosomes with the immunesystem. Philos Trans R Soc tond B Biol Sci. 1984 Nov. 13;307(1131):41-9; Allison A C, Eugui E M. The role of cell-mediated immuneresponses in resistance to malaria, with special reference to oxidantstress. Annu Rev Immunol. 1983; 1:361-92.)

NK cells have been shown to play a role in attacking cancer cells thatpresent membrane bound Hsp70. It is believed that membrane bound Hsp70binds to CD94 receptors on the surface of NK cells and cause them toproduce and secrete high amounts of the enzyme, granzyme B which isthought to enter the tumor cell via interaction with membrane boundHsp70 and induce apoptosis (see Radons and Multhoff, Exerc. Immunol.Rev. (2005), 11:17-33). Therefore, there is an urgent need for effectivetreatments for increasing NK cell activity for the treatment of cancerand other disorders that respond to NK induction.

SUMMARY OF THE INVENTION

Certain compounds of the invention induce Hsp70 production in cells andthereby increase the level of Hsp70 in the cytoplasm and on the surfaceof cells. In addition, certain compounds of the invention are cytotoxicto cancer cell lines, including multi-drug resistant cancer cell lines,and enhance the anti-proliferative and apoptotic activity (e.g.,anti-cancer activity) of Taxol and taxane analogs.

-   -   In one embodiment, the compounds of the invention are        represented by formula (I):

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X₁ is C(R₉)₂ or N—X₂;

X₂ is R₁₇, —OR₁₇, —N(R₂₆)₂, or —NHR₂₇;

Z₁, Z₂, Z₃, and Z₄ are each independently O or S;

R₁ and R₂ are independently an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, halo, nitro,cyano, guanidino, —OR₁₇, —NR₁₉R₂₀, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇,—C(O)NR₁₉R₂₀; —NR₁₈C(O)R₁₇, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇,—S(O)_(p)R₁₇, —OS(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₂, or—S(O)_(p)NR₁₉R₂₀;

R₃ and R₄ are independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl or an optionally substituted heteroaryl;

R₅ and R₆ are independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl or an optionally substituted heteroaryl;

each R₉ is independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, halo, nitro,cyano, guanidino, —OR₁₇, —NR₄OR₄₁, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇,—C(O)NR₁₉R₂₀, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇, —S(O)_(p)R₁₇,—S(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or —S(O)_(p)NR₁₉R₂₀;

R₁₇ and R₁₈, for each occurrence, are, independently, —H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl;

R₁₉ and R₂₀, for each occurrence, are independently —H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; or R₁₉ and R₂₀, taken together with the nitrogen to whichthey are attached, form an optionally substituted heterocyclyl or anoptionally substituted heteroaryl;

R₂₆ is an optionally substituted alkyl, an optionally substitutedalkenyl, an optionally substituted alkynyl, an optionally substitutedcycloalkyl, an optionally substituted cycloalkenyl, an optionallysubstituted heterocyclyl, an optionally substituted aryl, an optionallysubstituted heteroaryl, an optionally substituted aralkyl, or anoptionally substituted heteraralkyl;

R₂₇ is —H, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl, providedthat R₂₇ is not an optionally substituted phenyl;

R₄₀ and R₄₁, for each occurrence, are independently H, —OR₁₇, —NR₁₉R₂₀,—C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, anoptionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, or an optionallysubstituted heteraralkyl, or R₄₀ and R₄₁, taken together with thenitrogen to which they are attached, form an optionally substitutedheterocyclyl or an optionally substituted heteroaryl; and

p is 1 or 2.

In one embodiment of compounds of formula (I), the compound is notpropanedioic acid, (2-methylpropylidene)-,bis[2-[(phenylamino)thioxomethyl]hydrazide].

-   -   In another embodiment, the compounds of the invention are        represented by formula (II):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (III):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (IV):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

In another embodiment, the compounds of the invention are represented byformula (V):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (VI):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (VII):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

Another embodiment of the present invention is a pharmaceuticalcomposition comprising a compound of the invention, or a tautomer,pharmaceutically acceptable salt, solvate, clathrate, or a prodrugthereof, and a pharmaceutically acceptable carrier or diluent. Thepharmaceutical compositions can be used in therapy, for example, asanti-proliferative agents (e.g., anti-cancer agents). In addition, thepharmaceutical compositions can be used in therapy to treat disordersresponse to Hsp70 induction, such as cystic fibrosis, or thepharmaceutical compositions can be used in therapy to treat disordersresponse to natural killer cell induction, such as bacterial infections,fungal infections, viral infections, or parasitic infections.

The present invention also provides for a method of treating a subjectwith a proliferative disorder, such as cancer. The method comprisesadministering to the subject an effective amount of a compound of theinvention, or a tautomer, pharmaceutically acceptable salt, solvate,clathrate, or a prodrug thereof. The compound of the invention, or atautomer, pharmaceutically acceptable salt, solvate, clathrate, or aprodrug thereof, may be administered as a mono-therapy (i.e., as theonly anti-proliferative drug administered to the subject) or isco-administered with one or more other anti-cancer drugs. In oneembodiment, the compound of the invention, or a tautomer,pharmaceutically acceptable salt, solvate, clathrate, or a prodrugthereof, is administered with Taxol® or a taxane derivative.

The use of a compound of the invention, or a tautomer, pharmaceuticallyacceptable salt, solvate, clathrate, or a prodrug thereof, in themanufacture of a medicament for the purpose of treating a proliferativedisorder, such as cancer, in an individual is also provided in thepresent invention.

The present invention also provides for a method of treating a subjecthaving an Hsp70 responsive disorder, such as Alzheimer's disease,Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosisand cystic fibrosis. The method comprises administering to the subjectan effective amount of a compound of the invention, or a tautomer,pharmaceutically acceptable salt, solvate, clathrate, or a prodrugthereof.

The use of a compound of the invention, or a tautomer, pharmaceuticallyacceptable salt, solvate, clathrate, or a prodrug thereof, in themanufacture of a medicament for the purpose of disorders responsive toHsp70 induction, such as Alzheimer's disease, Huntington's disease,Parkinson's disease, amyotrophic lateral sclerosis and cystic fibrosis,in an individual is also provided in the present invention.

The present invention also provides for a method of treating a subjecthaving a natural killer cell responsive disorder, such as bacterialinfections, fungal infections, viral infections, or parasiticinfections. The method comprises administering to the subject aneffective amount of a compound of the invention, or a tautomer,pharmaceutically acceptable salt, solvate, clathrate, or a prodrugthereof.

The use of a compound of the invention, or a tautomer, pharmaceuticallyacceptable salt, solvate, clathrate, or a prodrug thereof, in themanufacture of a medicament for the purpose of disorders responsive tonatural killer cell induction, such as bacterial infection, fungalinfection, viral infection, or parasitic infections, in an individual isalso provided in the present invention.

The compounds of the invention or tautomer, pharmaceutically acceptablesalt, solvate, clathrate, or a prodrug thereof, can be used to treatproliferative disorders such as cancer, including cancers that havebecome multi-drug resistant, alone or in combination with otheranti-cancer agents. Thus, the compounds of the invention can be used totreat cancers where other drug regimens have either failed or becomeineffective. Additionally, the compounds of the invention areparticularly effective when used in combination with other anti-cancerdrugs such as Taxol or a taxane analog.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is the structure of Taxol® (paclitaxel).

FIG. 2 is the structure of Taxotere® (docetaxel).

FIGS. 3-23 each depict the structure of particular Taxol® analogs.

FIG. 24 is the structure of a polymer comprising a Taxol® analog grouppendent from the polymer backbone. The polymer is a terpolymer of thethree monomer units shown.

DETAILED DESCRIPTION OF THE INVENTION

-   -   In one embodiment, the compounds of the invention are        represented by formula (I):

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

X₁ is C(R₉)₂ or N—X₂;

X₂ is R₁₇, —OR₁₇, —N(R₂₆)₂, or —NHR₂₇;

Z₁, Z₂, Z₃, and Z₄ are each independently O or S;

R₁ and R₂ are independently an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, halo, nitro,cyano, guanidino, —OR₁₇, —NR₁₉R₂₀, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇,—C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, SR₁₇,—S(O)_(p)R₁₇, —OS(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or—S(O)_(p)NR₁₉R₂₀;

R₃ and R₄ are independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl or an optionally substituted heteroaryl;

R₅ and R₆ are independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl or an optionally substituted heteroaryl;

each R₉ is independently —H, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, halo, nitro,cyano, guanidino, —OR₁₇, —NR₄₀R₄₁, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇,—C(O)NR₁₉R₂₀, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇, —S(O)_(p)R₁₇,—OS(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or —S(O)_(p)NR₁₉R₂₀;

R₁₇ and R₁₈, for each occurrence, are, independently, —H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl;

R₁₉ and R₂₀, for each occurrence, are independently —H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; or R₁₉ and R₂₀, taken together with the nitrogen to whichthey are attached, form an optionally substituted heterocyclyl or anoptionally substituted heteroaryl;

R₂₆ is an optionally substituted alkyl, an optionally substitutedalkenyl, an optionally substituted alkynyl, an optionally substitutedcycloalkyl, an optionally substituted cycloalkenyl, an optionallysubstituted heterocyclyl, an optionally substituted aryl, an optionallysubstituted heteroaryl, an optionally substituted aralkyl, or anoptionally substituted heteraralkyl;

R₂₇ is —H, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl, providedthat R₂₇ is not an optionally substituted phenyl;

R₄₀ and R₄₁, for each occurrence, are independently H, —OR₁₇, —NR₁₉R₂₀,—C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, anoptionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, an optionally substituted aralkyl, or an optionallysubstituted heteraralkyl, or R₄₀ and R₄₁, taken together with thenitrogen to which they are attached, form an optionally substitutedheterocyclyl or an optionally substituted heteroaryl; and

p is 1 or 2.

In one embodiment of compounds of formula (I), the compound is notpropanedioic acid, (2-methylpropylidene)-,bis[2-[(phenylamino)thioxomethyl]hydrazide].

-   -   In another embodiment, the compounds of the invention are        represented by formula (II):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (III):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (IV):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

In another embodiment, the compounds of the invention are represented byformula (V):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (VI):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

-   -   In another embodiment, the compounds of the invention are        represented by formula (VII):

or a pharmaceutically acceptable salt or prodrug thereof, wherein thevariables are defined as for formula (I).

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are each independently an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, or an optionally substituted heteroaryl.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are each an optionally substituted aryl or an optionallysubstituted heteroaryl. In one aspect, R₁ and R₂ are each a substitutedor unsubstituted phenyl group. In one aspect, R₁ and R₂ are both phenyl.In another aspect, R₁ and R₂ are both 4-cyanophenyl. In another aspect,R₁ and R₂ are both 4-methoxyphenyl. In a further aspect, R₁ and R₂ areboth 2,5-dimethoxyphenyl. In another aspect, R₁ and R₂ are both3-cyanophenyl. In a further aspect, R₁ and R₂ are both 3-fluorophenyl.In another aspect, R₁ and R₂ are both 4-chlorophenyl. In a furtheraspect, R₁ and R₂ are both 2-dimethoxyphenyl. In another aspect, R₁ andR₂ are both 3-methoxyphenyl. In one aspect, R₁ and R₂ are both2,3-dimethoxyphenyl. In another aspect, R₁ and R₂ are both2,5-difluorophenyl. In a further aspect, R₁ and R₂ are both2,5-dichlorophenyl. In another aspect, R₁ and R₂ are both2,5-dimethylphenyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are both an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl. In one aspect, R₁ and R₂ are both aC₃-C₈ cycloalkyl group optionally substituted with at least one alkylgroup. In another aspect, R₁ and R₂ are both cyclopropyl or1-methylcyclopropyl. In one aspect, R₁ and R₂ are both cyclopropyl. In afurther aspect, R₁ and R₂ are both 1-methylcyclopropyl. In anotheraspect, R₁ and R₂ are both 2-methylcyclopropyl. In one aspect, R₁ and R₂are both 2-phenylcyclopropyl. In another aspect, R₁ and R₂ are both1-phenylcyclopropyl. In a further aspect, R₁ and R₂ are both cyclobutyl.In another aspect, R₁ and R₂ are both cyclopentyl. In another aspect, R₁and R₂ are both cyclohexyl. In a further aspect, R₁ and R₂ are bothmethyl. In another aspect, R₁ and R₂ are both methyl. In one aspect, R₁and R₂ are both t-butyl. In another aspect, R₁ and R₂ are ethyl. In afurther aspect, R₁ and R₂ are both n-propyl. In another aspect, R₁ andR₂ are haloalkyls.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are both halo, nitro, cyano, guanidino, —OR₁₇, —NR₁₉R₂₀,—C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇,—OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇, —S(O)_(p)R₁₇, —OS(O)_(p)R₁₇,—S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or —S(O)_(p)NR₁₉R₂₀. In one aspect, R₁and R₂ are both —OR₁₇. In one aspect, R₁ and R₂ are both —NR₁₉R₂₀.

In another embodiment of the compounds represented by formula (I)-(VI),R₃ and R₄ are each an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl. In one aspect, R₃ and R₄ are eachan alkyl group. In another aspect, R₃ and R₄ are each methyl or ethyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₅ is —H and R₆ is —H, an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl. In one aspect, R₆ is —H or methyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are each an optionally substituted aryl or an optionallysubstituted heteroaryl; and R₃ and R₄ are each an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₅ is —H and R₆ is —H, an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl; and R₃ and R₄ are each an alkylgroup.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are each a substituted or unsubstituted phenyl group and R₃and R₄ are each methyl or ethyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are both an optionally substituted alkyl, an optionallysubstituted alkenyl, an optionally substituted alkynyl, an optionallysubstituted cycloalkyl, an optionally substituted cycloalkenyl, anoptionally substituted heterocyclyl; R₅ is —H; and R₆ is —H or anoptionally substituted alkyl, an optionally substituted alkenyl, anoptionally substituted alkynyl, an optionally substituted cycloalkyl, anoptionally substituted cycloalkenyl, an optionally substitutedheterocyclyl.

In another embodiment of the compounds represented by formula (I)-(VI),R₁ and R₂ are both a C₃-C₈ cycloalkyl group optionally substituted withat least one alkyl group; R₃ and R₄ are both an alkyl group; and R₆ is—H or methyl.

In another embodiment of the compounds represented by formula (I)-(VI),

R₁ and R₂ are both phenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both phenyl; R₃ and R₄ are both ethyl;R₁ and R₂ are both 4-cyanophenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 4-methoxyphenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2,5-dimethoxyphenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 3-fluorophenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 4-chlorophenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2-dimethoxyphenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 3-methoxyphenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2,3-dimethoxyphenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2,5-difluorophenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2,5-dichlorophenyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclopropyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclopropyl; R₃ and R₄ are both ethyl;R₁ and R₂ are both 1-methylcyclopropyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 1-methylcyclopropyl; R₃ and R₄ are both ethyl;R₁ and R₂ are both 1-methylcyclopropyl; R₃ is methyl, and R₄ is ethyl;R₁ and R₂ are both 2-methylcyclopropyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 2-phenylcyclopropyl; R₃ and R₄ are both methyl;R₁ and R₂ are both 1-phenylcyclopropyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclobutyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclopentyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclohexyl; R₃ and R₄ are both methyl;R₁ and R₂ are both cyclohexyl; R₃ and R₄ are both phenyl;R₁ and R₂ are both methyl; R₃ and R₄ are both methyl;R₁ and R₂ are both methyl; R₃ and R₄ are both t-butyl;R₁ and R₂ are both methyl; R₃ and R₄ are both phenyl;R₁ and R₂ are both t-butyl; R₃ and R₄ are both methyl;R₁ and R₂ are ethyl; R₃ and R₄ are both methyl; orR₁ and R₂ are both n-propyl; R₃ and R₄ are both methyl.

In another embodiment of the compounds represented by formula (I),(III), or (V), Z₁ and Z₂ are both I.

In another embodiment of the compounds represented by formula (I),(III), or (V), Z₁ and Z₂ are both S.

In another embodiment of the compounds represented by formula (I),(III), or (V), Z₃ and Z₄ are both O.

In another embodiment of the compounds represented by formula (I),(III), or (V), Z₃ and Z₄ are both S.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₁ is C(R₉)₂.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₁ is N—X₂.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is R₁₇, —OR₁₇, or —N(R₂₆)₂.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is R₁₇.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is —OR₁₇.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is —N(R₂₆)₂.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is —NHR₂₇.

In another embodiment of the compounds represented by formula (I), (II),or (VII), X₂ is —OR₁₇, —N(R₂₆)₂ or —NHR₂₇.

In another embodiment of the compounds represented by formula (I), (II),or (VII), each R₉ is independently —H, an optionally substituted alkyl,an optionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, or —NR₄OR₄₁. Inone aspect, each R₉ is independently —H, an optionally substitutedalkyl, an optionally substituted cycloalkyl, an optionally substitutedheterocyclyl, an optionally substituted aryl, an optionally substitutedheteroaryl, or —NR₄₀R₄₁. In one aspect, each R₉ is independently —H, anoptionally substituted alkyl, an optionally substituted cycloalkyl, or—NR₄₀R₄₁. In one aspect, each R₉ is independently —H or —NR₄₀R₄₁.

In another embodiment of the compounds represented by formula (I), (II),(V), (VI), or (VII), R₁₇ is —H or an optionally substituted alkyl. Inone aspect, R₁₇ is —H or an optionally substituted C₁-C₆ alkyl. In oneaspect, R₁₇ is —H.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), or (VII), R₄₀ and R₄₁, for each occurrence, areindependently H, —OR₁₇, —NR₁₉R₂₀, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇,—CO(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, an optionally substituted alkyl, anoptionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl. In oneaspect, R₄₀ and R₄₁, for each occurrence, —H, —NR₁₉R₂₀, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl. In one aspect, R₄₀ and R₄₁, for each occurrence, areindependently —H, an optionally substituted alkyl, —NR₁₉R₂₀, anoptionally substituted cycloalkyl, or an optionally substitutedheterocyclyl. In one aspect, R₄₀ and R₄₁, for each occurrence, areindependently —H, an optionally substituted C₁-C₆ alkyl, —NR₁₉R₂₀, anoptionally substituted C3-C6 cycloalkyl, or an optionally substitutedC5-C6 heterocyclyl.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), or (VII), R₄₀ and R₄₁, taken together with the nitrogen towhich they are attached, form an optionally substituted heterocyclyl oran optionally substituted heteroaryl.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), or (VII), when R₄₀ and R₄₁ are —NR₁₉R₂₀, R₁₉ and R₂₀ areindependently —H or optionally substituted alkyl. In one aspect, R₁₉ andR₂₀ are independently —H or optionally substituted alkyl, wherein thealkyl is optionally substituted with —OH or —OC1-C6 alkyl.

In another embodiment of the compounds represented by formula (I), (II),(III), (IV), or (VII), R₄₀ and R₄₁ are independently —H, methyl, ethyl,—C1-C4alkylOH, —C1-C4alkylOC1-C4 alkyl, —OC1-C4 alkyl, dimethylamino,cyclopropyl, morpholinyl, or piperazinyl.

In another embodiment, the compound is selected from the groupconsisting of

-   2-((dimethylamino)methylene)-N′ 1,N′3-dimethyl-N′    1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-(hydroxyimino)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-(methoxyimino)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-((2-hydroxyethylamino)methylene)-N′1,N′3-dimethyl-N′    1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-((2-methoxyethylamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-((methoxyamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-((2,2-dimethylhydrazinyl)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-(aminomethylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   N′1,N′3-dimethyl-2-((methylamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   2-((cyclopropylamino)methylene)-N′    1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;-   N′1,N′3-dimethyl-2-((morpholinoamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide;    or-   N′ 1,N′3-dimethyl-2-((4-methylpiperazin-1-ylamino)methylene)-N′    1,N′3-di(phenylcarbonothioyl)malonohydrazide;

or a pharmaceutically acceptable salt or prodrug thereof, or apharmaceutically acceptable salt or prodrug thereof.

Exemplary compounds of the invention are depicted in Table 1 below,including tautomers, pharmaceutically acceptable salts, solvates,clathrates, hydrates, polymorphs or prodrugs thereof

TABLE 1 Compound Number Structure Name 1

2-((dimethylamino)methylene)- N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl) malonohydrazide 2

2-(hydroxyimino)-N′1,N′3- dimethyl-N′1,N′3- di(phenylcarbonothioyl)malonohydrazide 3

2-(methoxyimino)-N′1,N′3- dimethyl-N′1,N′3- di(phenylcarbonothioyl)malonohydrazide 4

2-((2- hydroxyethylamino)methylene)- N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 5

2-((2- methoxyethylamino)methylene)- N′1,N′3-dimethyl-N′1,N′3-(phenylcarbonothioyl)malonohydrazide 6

2-((methoxyamino)methylene)- N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 7

2-((2,2- dimethylhydrazinyl)methylene)- N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 8

2-(aminomethylene)-N′1,N′3- dimethyl-N′1,N′3- di(phenylcarbonothioyl)malonohydrazide 9

N′1,N′3-dimethyl-2- ((methylamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 10

2-((cyclopropylamino)methylene)- N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 11

N′1,N′3-dimethyl-2- ((morpholinoamino)methylene)- N′1,N′3-di(phenylcarbonothioyl)malonohydrazide 12

N′1,N′3-dimethyl-2-((4- methylpiperazin-1- ylamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

In one embodiment, the compounds of the invention do not include thecompounds disclosed in the patents and patent applications listed inTable 2.

TABLE 2 Patent or Patent Publication Application No. Publication No.Filing Date Date U.S. Pat. No. U.S. 2005/0009920 Jul. 10, 2002 Jun. 26,2003 6,800,660 U.S. Pat. No. U.S. 2003/0119914 May 14, 2004 Jan. 13,2005 7,037,940 U.S. Pat. No. U.S. 2006/0122183 Oct. 5, 2005 Jun. 8, 200611/244,324 U.S. Pat. No. U.S. 2003/0045518 Jul. 10, 2002 Mar. 6, 20036,762,204 U.S. Pat. No. U.S. 2003/0195258 Jan. 15, Oct. 16, 20036,924,312 2003 U.S. Pat. No. U.S. 2004/0235909 Mar. 18, 2004 Nov. 25,2004 7,001,923 U.S. Pat. No. U.S. 2006/0116374 Oct. 5, 2005 Jun. 1, 200611/244,427 U.S. Pat. No. U.S. 2003/0069225 Jul. 10, 2002 Apr. 10, 20036,825,235 U.S. Pat. No. U.S. 2004/0229952 Mar. 24, 2004 Nov. 18, 20047,074,952 U.S. Pat. No. Not published May 24, 2006 Not published11/440,429 U.S. Pat. No. U.S. 2006/0135595 Jun. 20, 2005 Jun. 22, 200611/157,213 U.S. Pat. No. Not published May 11, 2006 Not published11/432,307

As used herein, the term “alkyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 1 to 10 carbon atoms.Representative saturated straight chain alkyls include methyl, ethyl,n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl andn-decyl; while saturated branched alkyls include isopropyl, sec-butyl,isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl,2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl,3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl,2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl,2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl,2,2-dimethylhexyl, 3,3-dimtheylpentyl, 3,3-dimethylhexyl,4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl,3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl,2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl,2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl,3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. Theterm “(C₁-C₆)alkyl” means a saturated straight chain or branchednon-cyclic hydrocarbon having from 1 to 6 carbon atoms. Representative(C₁-C₆)alkyl groups are those shown above having from 1 to 6 carbonatoms. Alkyl groups included in compounds of this invention may beoptionally substituted with one or more substituents.

As used herein, the term “alkenyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 2 to 10 carbon atoms andhaving at least one carbon-carbon double bond. Representative straightchain and branched (C₂-C₁₀)alkenyls include vinyl, allyl, 1-butenyl,2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl,3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl,3-decenyl and the like. Alkenyl groups may be optionally substitutedwith one or more substituents.

As used herein, the term “alkynyl” means a saturated straight chain orbranched non-cyclic hydrocarbon having from 2 to 10 carbon atoms andhaving at lease one carbon-carbon triple bond. Representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, 4-pentynyl,1-hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl,1-octynyl, 2-octynyl, 7-octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl,1-decynyl, 2-decynyl, 9-decynyl, and the like. Alkynyl groups may beoptionally substituted with one or more substituents.

As used herein, the term “cycloalkyl” means a saturated, mono- orpolycyclic alkyl radical having from 3 to 20 carbon atoms.Representative cycloalkyls include cyclopropyl, 1-methylcyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,cyclononyl, -cyclodecyl, octahydro-pentalenyl, and the like. Cycloalkylgroups may be optionally substituted with one or more substituents.

As used herein, the term “cycloalkenyl” means a mono- or poly-cyclicnon-aromatic alkyl radical having at least one carbon-carbon double bondin the cyclic system and from 3 to 20 carbon atoms. Representativecycloalkenyls include cyclopentenyl, cyclopentadienyl, cyclohexenyl,cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl,cyclooctenyl, cyclooctadienyl, cyclooctatrienyl, cyclooctatetraenyl,cyclononenyl, cyclononadienyl, cyclodecenyl, cyclodecadienyl,1,2,3,4,5,8-hexahydronaphthalenyl and the like. Cycloalkenyl groups maybe optionally substituted with one or more substituents.

As used herein, the term “haloalkyl” means and alkyl group in which oneor more (including all) the hydrogen radicals are replaced by a halogroup, wherein each halo group is independently selected from —F, —Cl,—Br, and —I. The term “halomethyl” means a methyl in which one to threehydrogen radical(s) have been replaced by a halo group. Representativehaloalkyl groups include trifluoromethyl, bromomethyl,1,2-dichloroethyl, 4-iodobutyl, 2-fluoropentyl, and the like.

As used herein, an “alkoxy” is an alkyl group which is attached toanother moiety via an oxygen linker.

As used herein, a “haloalkoxy” is a haloalkyl group which is attached toanother moiety via an oxygen linker.

As used herein, the term an “aromatic ring” or “aryl” means ahydrocarbon monocyclic or polycyclic radical in which at least one ringis aromatic. Examples of suitable aryl groups include, but are notlimited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl,and naphthyl, as well as benzo-fused carbocyclic moieties such as5,6,7,8-tetrahydronaphthyl. Aryl groups may be optionally substitutedwith one or more substituents. In one embodiment, the aryl group is amonocyclic ring, wherein the ring comprises 6 carbon atoms, referred toherein as “(C₆)aryl.”

As used herein, the term “aralkyl” means an aryl group that is attachedto another group by a (C₁-C₆)alkylene group. Representative aralkylgroups include benzyl, 2-phenyl-ethyl, naphth-3-yl-methyl and the like.Aralkyl groups may be optionally substituted with one or moresubstituents.

As used herein, the term “alkylene” refers to an alkyl group that hastwo points of attachment. The term “(C₁-C₆)alkylene” refers to analkylene group that has from one to six carbon atoms. Straight chain(C₁-C₆)alkylene groups are preferred. Non-limiting examples of alkylenegroups include methylene (—CH₂—), ethylene (—CH₂CH₂—), n-propylene(—CH₂CH₂CH₂—), isopropylene (—CH₂CH(CH₃)—), and the like. Alkylenegroups may be optionally substituted with one or more substituents.

As used herein, the term “heterocyclyl” means a monocyclic (typicallyhaving 3- to 10-members) or a polycyclic (typically having 7- to20-members) heterocyclic ring system which is either a saturated ring oran unsaturated non-aromatic ring. A 3- to 10-membered heterocycle cancontain up to 5 heteroatoms; and a 7- to 20-membered heterocycle cancontain up to 7 heteroatoms. Typically, a heterocycle has at least oncarbon atom ring member. Each heteroatom is independently selected fromnitrogen, which can be oxidized (e.g., N(O)) or quaternized; oxygen; andsulfur, including sulfoxide and sulfone. The heterocycle may be attachedvia any heteroatom or carbon atom. Representative heterocycles includemorpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl,tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, andthe like. A heteroatom may be substituted with a protecting group knownto those of ordinary skill in the art, for example, the hydrogen on anitrogen may be substituted with a tert-butoxycarbonyl group.Furthermore, the heterocyclyl may be optionally substituted with one ormore substituents. Only stable isomers of such substituted heterocyclicgroups are contemplated in this definition.

As used herein, the term “heteroaromatic”, “heteroaryl” or like termsmeans a monocyclic or polycyclic heteroaromatic ring comprising carbonatom ring members and one or more heteroatom ring members. Eachheteroatom is independently selected from nitrogen, which can beoxidized (e.g., N(O)) or quaternized; oxygen; and sulfur, includingsulfoxide and sulfone. Representative heteroaryl groups include pyridyl,1-oxo-pyridyl, furanyl, benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl,pyrrolyl, oxazolyl, imidazolyl, thiazolyl, a isoxazolyl, quinolinyl,pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, atriazinyl, triazolyl, thiadiazolyl, isoquinolinyl, indazolyl,benzoxazolyl, benzofuryl, indolizinyl, imidazopyridyl, tetrazolyl,benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl,indolyl, tetrahydroindolyl, azaindolyl, imidazopyridyl, quinazolinyl,purinyl, pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl,imidazo[1,2-a]pyridyl, and benzothienyl. In one embodiment, theheteroaromatic ring is selected from 5-8 membered monocyclic heteroarylrings. The point of attachment of a heteroaromatic or heteroaryl ring toanother group may be at either a carbon atom or a heteroatom of theheteroaromatic or heteroaryl rings. Heteroaryl groups may be optionallysubstituted with one or more substituents.

As used herein, the term “(C₅)heteroaryl” means an aromatic ring of 5members, wherein at least one atom in the ring is a heteroatom such as,for example, oxygen, sulfur or nitrogen. Representative (C₅)heteroarylsinclude furanyl, thienyl, pyrrolyl, oxazolyl, imidazolyl, thiazolyl,isoxazolyl, pyrazolyl, isothiazolyl, pyrazinyl, triazolyl, thiadiazolyl,and the like.

As used herein, the term “(C₆)heteroaryl” means an aromatic ring of 6members, wherein at least one atom in the ring is a heteroatom such as,for example, oxygen, nitrogen or sulfur. Representative (C₆)heteroarylsinclude pyridyl, pyridazinyl, pyrazinyl, triazinyl, tetrazinyl and thelike.

As used herein, the term “heteroaralkyl” means a heteroaryl group thatis attached to another group by a (C₁-C₆)alkylene. Representativeheteroaralkyls include 2-(pyridin-4-yl)-propyl, 2-(thien-3-yl)-ethyl,imidazol-4-yl-methyl and the like. Heteroaralkyl groups may beoptionally substituted with one or more substituents.

As used herein, the term “halogen” or “halo” means —F, —Cl, —Br or —I.

Suitable substituents for an alkyl, alkylene, alkenyl, alkynyl,cycloalkyl, cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, andheteroaralkyl groups include any substituent which will form a stablecompound of the invention. Examples of substituents for an alkyl,alkylene, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocyclyl,aryl, aralkyl, heteroaryl, and heteroarylalkyl include an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, an optionally substituted heteraralkyl,a haloalkyl, —C(O)NR₂₈R₂₉, —C(S)NR₂₈R₂₉, —C(NR₃₂)NR₂₈R₂₉, —NR₃₀C(O)R₃₁,—NR₃₀C(S)R₃₁, —NR₃₀C(NR₃₂)R₃₁, halo, —OR₃₀, cyano, nitro, haloalkoxy,—C(O)R₃₀, —C(S)R₃₀, —C(NR₃₂)R₃₀, —NR₂₈R₂₉, —C(O)OR₃₀, —C(S)OR₃₀,—C(NR₃₂)OR₃₀, —OC(O)R₃₀, —OC(S)R₃₀, —OC(NR₃₂)R₃₀, —NR₃₀C(O)NR₂₈R₂₉,—NR₃₀C(S)NR₂₈R₂₉, —NR₃₀C(NR₃₂)NR₂₈R₂₉, —OC(O)NR₂₈R₂₉, —OC(S)NR₂₈R₂₉,—OC(NR₃₂)NR₂₈R₂₉, —NR₃₀C(O)OR₃₁, —NR₃₀C(S)OR₃₁, —NR₃₀C(NR₃₂)OR₃₁,—S(O)_(h)R₃₀, —OS(O)_(p)R₃₀, —NR₃₀S(O)_(p)R₃₀, —S(O)_(p)NR₂₈R₂₉,—OS(O)_(p)NR₂₈R₂₉, or —NR₃₀S(O)_(p)NR₂₈R₂₉, wherein R₂₈ and R₂₉, foreach occurrence are, independently, H, an optionally substituted alkyl,an optionally substituted alkenyl, an optionally substituted alkynyl, anoptionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, or an optionally substituted heteraralkyl; or R₂₈and R₂₉ taken together with the nitrogen to which they are attached isoptionally substituted heterocyclyl or optionally substitutedheteroaryl.

R₃₀ and R₃₁ for each occurrence are, independently, H, an optionallysubstituted alkyl, an optionally substituted alkenyl, an optionallysubstituted alkynyl, an optionally substituted cycloalkyl, an optionallysubstituted cycloalkenyl, an optionally substituted heterocyclyl, anoptionally substituted aryl, an optionally substituted heteroaryl, anoptionally substituted aralkyl, or an optionally substitutedheteraralkyl; and

R₃₂, for each occurrence is, independently, H, an optionally substitutedalkyl, an optionally substituted alkenyl, an optionally substitutedalkynyl, an optionally substituted cycloalkyl, an optionally substitutedcycloalkenyl, an optionally substituted heterocyclyl, an optionallysubstituted aryl, an optionally substituted heteroaryl, an optionallysubstituted aralkyl, an optionally substituted heteraralkyl, —C(O)R₃₀,—C(O)NR₂₈R₂₉, —S(O)_(p)R₃₀, or —S(O)_(p)NR₂₈R₂₉;

p is 1 or 2; and

h is 0, 1 or 2.

In addition, alkyl, cycloalkyl, alkylene, a heterocyclyl, and anysaturated portion of a alkenyl, cycloalkenyl, alkynyl, aralkyl, andheteroaralkyl groups, may also be substituted with ═O, ═S, ═N—R₃₂.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

As used herein, the terms “subject”, “patient” and “mammal” are usedinterchangeably. The terms “subject” and “patient” refer to an animal(e.g., a bird such as a chicken, quail or turkey, or a mammal),preferably a mammal including a non-primate (e.g., a cow, pig, horse,sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and a primate(e.g., a monkey, chimpanzee and a human), and more preferably a human.In one embodiment, the subject is a non-human animal such as a farmanimal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog, cat,guinea pig or rabbit). In a preferred embodiment, the subject is ahuman.

As used herein, the term “lower” refers to a group having up to fourcarbon atoms. For example, a “lower alkyl” refers to an alkyl radicalhaving from 1 to 4 carbon atoms, “lower alkoxy” refers to“—O—(C₁-C₄)alkyl and a “lower alkenyl” or “lower alkynyl” refers to analkenyl or alkynyl radical having from 2 to 4 carbon atoms,respectively.

Unless indicated otherwise, the compounds of the invention containingreactive functional groups (such as (without limitation) carboxy,hydroxy, thiol, and amino moieties) also include protected derivativesthereof. “Protected derivatives” are those compounds in which a reactivesite or sites are blocked with one or more protecting groups. Examplesof suitable protecting groups for hydroxyl groups include benzyl,methoxymethyl, allyl, trimethylsilyl, tert-butyldimethylsilyl, acetate,and the like. Examples of suitable amine protecting groups includebenzyloxycarbonyl, tert-butoxycarbonyl, tert-butyl, benzyl andfluorenylmethyloxy-carbonyl (Fmoc). Examples of suitable thiolprotecting groups include benzyl, tert-butyl, acetyl, methoxymethyl andthe like. Other suitable protecting groups are well known to those ofordinary skill in the art and include those found in T. W. Greene,Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981.

As used herein, the term “compound(s) of this invention” and similarterms refers to a compound of formula (I) through (VII) and Table 1, ora pharmaceutically acceptable salt, solvate, clathrate, hydrate,polymorph or prodrug thereof, and also include protected derivativesthereof.

When a disclosed compound is named or depicted by structure, it is to beunderstood that solvates (e.g., hydrates) of the compound or itspharmaceutically acceptable salts are also included. “Solvates” refer tocrystalline forms wherein solvent molecules are incorporated into thecrystal lattice during crystallization. Solvate may include water ornonaqueous solvents such as ethanol, isopropanol, DMSO, acetic acid,ethanolamine, and EtOAc. Solvates, wherein water is the solvent moleculeincorporated into the crystal lattice, are typically referred to as“hydrates”. Hydrates include a stoichiometric or non-stoichiometricamount of water bound by non-covalent intermolecular forces.

When a disclosed compound is named or depicted by structure, it is to beunderstood that the compound, including solvates thereof, may exist incrystalline forms, non-crystalline forms or a mixture thereof. Thecompounds or solvates may also exhibit polymorphism (i.e. the capacityto occur in different crystalline forms). These different crystallineforms are typically known as “polymorphs.” It is to be understood thatwhen named or depicted by structure, the disclosed compounds andsolvates (e.g., hydrates) also include all polymorphs thereof. As usedherein, the term “polymorph” means solid crystalline forms of a compoundof the present invention or complex thereof. Different polymorphs of thesame compound can exhibit different physical, chemical and/orspectroscopic properties. Different physical properties include, but arenot limited to stability (e.g., to heat or light), compressibility anddensity (important in formulation and product manufacturing), anddissolution rates (which can affect bioavailability). Differences instability can result from changes in chemical reactivity (e.g.,differential oxidation, such that a dosage form discolors more rapidlywhen comprised of one polymorph than when comprised of anotherpolymorph) or mechanical characteristics (e.g., tablets crumble onstorage as a kinetically favored polymorph converts to thermodynamicallymore stable polymorph) or both (e.g., tablets of one polymorph are moresusceptible to breakdown at high humidity). Different physicalproperties of polymorphs can affect their processing. For example, onepolymorph might be more likely to form solvates or might be moredifficult to filter or wash free of impurities than another due to, forexample, the shape or size distribution of particles of it. In addition,one polymorph may spontaneously convert to another polymorph undercertain conditions.

When a disclosed compound is named or depicted by structure, it is to beunderstood that clathrates (“inclusion compounds”) of the compound orits pharmaceutically acceptable salts, solvates or polymorphs are alsoincluded. As used herein, the term “clathrate” means a compound of thepresent invention or a salt thereof in the form of a crystal latticethat contains spaces (e.g., channels) that have a guest molecule (e.g.,a solvent or water) trapped within.

The compounds of the invention may contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers, ordiastereomers. According to this invention, the chemical structuresdepicted herein, including the compounds of this invention, encompassall of the corresponding compounds' enantiomers, diastereomers andgeometric isomers, that is, both the stereochemically pure form (e.g.,geometrically pure, enantiomerically pure, or diastereomerically pure)and isomeric mixtures (e.g., enantiomeric, diastereomeric and geometricisomeric mixtures). In some cases, one enantiomer, diastereomer orgeometric isomer will possess superior activity or an improved toxicityor kinetic profile compared to other isomers. In those cases, suchenantiomers, diastereomers and geometric isomers of compounds of thisinvention are preferred.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may become active upon suchreaction under biological conditions, or they may have activity in theirunreacted forms. Examples of prodrugs contemplated in this inventioninclude, but are not limited to, analogs or derivatives of compounds offormula (I) through (VII) and Table 1 that comprise biohydrolyzablemoieties such as biohydrolyzable amides, biohydrolyzable esters,biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzableureides, and biohydrolyzable phosphate analogues. Other examples ofprodrugs include derivatives of compounds of formula (I) through (VII)and Table 1, that comprise —NO, —NO₂, —ONO, or —ONO₂ moieties. Prodrugscan typically be prepared using well-known methods, such as thosedescribed by 1 BURGER'S MEDICINAL CHEMISTRY AND DRUG DISCOVERY (1995)172-178, 949-982 (Manfred E. Wolff ed., 5^(th) ed).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide”, “biohydrolyzable ester”, “biohydrolyzablecarbamate”, “biohydrolyzable carbonate”, “biohydrolyzable ureide” and“biohydrolyzable phosphate analogue” mean an amide, ester, carbamate,carbonate, ureide, or phosphate analogue, respectively, that either: 1)does not destroy the biological activity of the compound and confersupon that compound advantageous properties in vivo, such as improvedwater solubility, improved circulating half-life in the blood (e.g.,because of reduced metabolism of the prodrug), improved uptake, improvedduration of action, or improved onset of action; or 2) is itselfbiologically inactive but is converted in vivo to a biologically activecompound. Examples of biohydrolyzable amides include, but are notlimited to, lower alkyl amides, α-amino acid amides, alkoxyacyl amides,and alkylaminoalkylcarbonyl amides. Examples of biohydrolyzable estersinclude, but are not limited to, lower alkyl esters, alkoxyacyloxyesters, alkyl acylamino alkyl esters, and choline esters. Examples ofbiohydrolyzable carbamates include, but are not limited to, loweralkylamines, substituted ethylenediamines, amino acids,hydroxyalkylamines, heterocyclic and heteroaromatic amines, andpolyether amines.

As used herein, “Hsp70” includes each member of the family of heat shockproteins having a mass of about 70-kiloDaltons, including forms such asconstitutive, cognate, cell-specific, glucose-regulated, inducible, etc.Examples of specific Hsp70 proteins include hsp70, hsp70hom; hsc70;Grp78/BiP; mt-hsp70/Grp75, and the like). Typically, the disclosedmethods increase expression of inducible Hsp70. Functionally, the 70-kDaHSP(HSP70) family is a group of chaperones that assist in the folding,transport, and assembly of proteins in the cytoplasm, mitochondria, andendoplasmic reticulum. Membrane-bound Hsp70 In humans, the Hsp70 familyencompasses at least 11 genes encoding a group of highly relatedproteins. See, for example, Tavaria, et al., Cell Stress Chaperones,1996; 1(1):23-28; Todryk, et al., Immunology. 2003, 110(1): 1-9; andGeorgopoulos and Welch, Annu Rev Cell Biol. 1993; 9:601-634; the entireteachings of these documents are incorporated herein by reference.

As used herein, an “Hsp70-responsive disorder” is a medical conditionwherein stressed cells can be treated by increased Hsp70 expression.Such disorders can be caused by a wide variety of cellular stressors,including, but not limited to Alzheimer's disease; Huntington's disease;Parkinson's disease; spinal/bulbar muscular atrophy (e.g., Kennedy'sdisease), spinocerebellar ataxic disorders, and other neuromuscularatrophies; familial amyotrophic lateral sclerosis; ischemia; seizure;hypothermia; hyperthermia; burn trauma; atherosclerosis; radiationexposure; glaucoma; toxin exposure; mechanical injury; inflammation;autoimmune disease; infection (bacterial, viral, fungal, or parasitic);and the like.

In some embodiments, the Hsp70-responsive disorder is aneurodegenerative disorder. As used herein, a neurodegenerative disorderinvolves degradation of neurons such as cerebral, spinal, and peripheralneurons (e.g., at neuromuscular junctions), more typically degradationof cerebral and spinal neurons, or in preferred embodiments, degradationof cerebral neurons. Neurodegenerative disorders can include Alzheimer'sdisease; Huntington's disease; Parkinson's disease; spinal/bulbarmuscular atrophy and other neuromuscular atrophies; and familialamyotrophic lateral sclerosis or other diseases associated withsuperoxide dismutase (SOD) mutations. Neurodegenerative disorders canalso include degradation of neurons caused by ischemia, seizure, thermalstress, radiation, toxin exposure, infection, injury, and the like.

In some embodiments, the Hsp70-responsive disorder is a disorder ofprotein aggregation/misfolding, such as Alzheimer's disease;Huntington's disease; Parkinson's disease; spongiform encephalopathies;and the like.

In another embodiment the Hsp70 responsive disorder is a treatment orcondition which causes or may cause nerve damage. The compounds for usein the methods of the present invention can be used to reduce or prevent(inhibit the onset of) nerve damage (i.e., provide neuroprotection) in asubject i) suffering from a condition which causes or may cause nervedamage or ii) receiving treatment which causes or may cause nervedamage. In one aspect, the treatment which causes or may cause nervedamage is radiation therapy. In another aspect, the treatment ischemotherapy. In one aspect, the chemotherapy comprises administering anantimitotic agent (e.g. vincristine, vinorelbine, paclitaxel, or apaclitaxel analog). In one aspect, the chemotherapy comprisesadministering paclitaxel. In another aspect, the chemotherapy comprisesadministering a platinum derivative (e.g. cisplatinum, carboplatin, oroxaliplatin). In certain embodiments, the compounds for use in themethods of the present invention can be administered simultaneously as acombination therapy with the treatment which causes or may cause nervedamage. In other embodiments the compounds for use in the methods of thepresent invention can be administered before or after the treatmentwhich causes may cause nerve damage. In certain embodiments thecompounds for use in the methods of the present invention can beadministered between 30 minutes and 12 hours, between 1 hour and 6before or after the treatment which causes or may cause nerve damage.

Nerve damage may be caused by a number of treatments including, but notlimited to, radiation therapy; chemotherapy, e.g. cisplatinum,carboplatin, oxaliplatin, vincristine, vinblastine, vinorelbine,vindesine, ifosfamide, methotrexate, cladribine, altretamine,fludarabine, procarbazine, thiotepa, teniposide, arsenic trioxide,alemtuzumab, capecitabine, dacarbazine, denileukin diftitox, interferonalpha, liposomal daunorubicin, tretinoin, etoposide/VP-16, cytarabine,hexamethylmelamine, suramin, paclitaxel, docetaxel, gemcitibine,thalidomide, and bortezomib; heart or blood pressure medications, e.g.amiodarone, hydralazine, digoxin, and perhxiline; medications to fightinfection, e.g. metronidazole, nitrofurantoin, thalidomide, and INH;medications to treat skin conditions, e.g. dapsone; anticonvulsants,e.g. phenyloin; anti-alcohol medications, e.g. disulfuram; HIVmedications, e.g. zidovudine, didanonsine, stavudine, zalcitabine,ritonavir, d4T, ddC, ddI, and amprenavir; cholesterol medications, e.g.lovastatin, pravastatin, indapamid, simvastatin, fluvastatin,atorvastatin, cerivastatin, and gemfibrozil; antirheumatics, e.g.chloroquine, cholchicine, organic gold, and penicillamine; nitrousoxide; lithium; and ergots.

In some embodiments, the Hsp70-responsive disorder is ischemia. Ischemiacan damage tissue through multiple routes, including oxygen depletion,glucose depletion, oxidative stress upon reperfusion, and/or glutamatetoxicity, and the like. Ischemia can result from an endogenous condition(e.g., stroke, heart attack, and the like), from accidental mechanicalinjury, from surgical injury (e.g., reperfusion stress on transplantedorgans), and the like. Alternatively, tissues that can be damaged byischemia include neurons, cardiac muscle, liver tissue, skeletal muscle,kidney tissue, pulmonary tissue, pancreatic tissue, and the like. In onepreferred embodiment, the Hsp70-responsive disorder is cerebral orspinal ischemia. In another preferred embodiment, the Hsp70-responsivedisorder is cardiac ischemia.

In various embodiments, the Hsp70-responsive disorder is seizure, e.g.,eplileptic seizure, injury-induced seizure, chemically-induced seizure,and the like.

In some embodiments, the Hsp70-responsive disorder is due to thermalstress. Thermal stress includes hyperthermia (e.g., from fever, heatstroke, burns, and the like) and hypothermia. In a preferred embodimentthe disorder is hyperthermia. In another preferred embodiment, theHsp70-responsive disorder is burn trauma.

In preferred embodiments, the Hsp70-responsive disorder isatherosclerosis.

In various embodiments, the Hsp70-responsive disorder is radiationdamage, e.g., due to visible light, ultraviolet light, microwaves,cosmic rays, alpha radiation, beta radiation, gamma radiation, X-rays,and the like. For example, the damage could be radiation damage tonon-cancerous tissue in a subject treated for cancer by radiationtherapy. In a preferred embodiment, the Hsp70-responsive disorder isradiation damage from visible light or ultraviolet light.

In various embodiments, the Hsp70-responsive disorder is mechanicalinjury, e.g., trauma from surgery, accidents, certain disease conditions(e.g., pressure damage in glaucoma) and the like. In a preferredembodiment, the Hsp70-responsive disorder is cerebral or spinal trauma.In another preferred embodiment, the Hsp70-responsive disorder isglaucoma (leading to pressure damage to retinal ganglions).

In various embodiments, the Hsp70-responsive disorder is exposure to atoxin. In preferred embodiments, the Hsp70-responsive disorder isexposure to a neurotoxin selected from methamphetamine; antiretroviralHIV therapeutics (e.g., nucleoside reverse transcriptase inhibitors;heavy metals (e.g., mercury, lead, arsenic, cadmium, compounds thereof,and the like), amino acid analogs, chemical oxidants, ethanol,glutamate, metabolic inhibitors, antibiotics, and the like.

Certain compounds of the invention also increase Natural Killer (NK)cell activity. As used herein, a “NK cell-responsive disorder” is amedical condition which is improved by an increased in NK cell activity.For example, a subject with a NK cell-responsive disorder may needimmune system augmentation because of infection or the possibilitythereof. In some embodiments, such a subject can have an infection (orhas been exposed to an infectious environment where pathogens arepresent, e.g., in a hospital) the symptoms of which may be alleviated bythe methods disclosed herein. For example, a subject in need oftreatment can have an infection (bacterial, viral, fungal, orparasitical (protozoal) for which the disclosed methods of activating NKcells can be a treatment.

In some embodiments, a subject having an NK cell-responsive disorder hasan immunodeficiency. Such a subject is in need of or can benefit fromprophylactic therapy, for example, a subject that has incomplete,damaged or otherwise compromised defenses against infection, or issubject to an infective environment, or the like. For example, a subjectcan be in an infectious environment where pathogens are present, e.g.,in a hospital; can have an open wound or burn injury; can have aninherited or acquired immune deficiency (e.g., severe combinedimmunodeficiency or “bubble boy” syndrome, variable immunodeficiencysyndrome acquired immune deficiency syndrome (AIDS), or the like); canhave a depressed immune system due to physical condition, age, toxinexposure, drug effect (immunosuppressants, e.g., in a transplantrecipient) or side effect (e.g., due to an anticancer agent); or thelike.

In some embodiments, NK cell activity can be increased in subjects thathave decreased or deficient NK cell activity, in conditions such aschronic fatigue syndrome (chronic fatigue immune dysfunction syndrome)or Epstein-Barr virus infection, post viral fatigue syndrome,post-transplantation syndrome (especially allogeneic transplants) orhost-graft disease, exposure to drugs such as anticancer agents ornitric oxide synthase inhibitors, natural aging, and variousimmunodeficient conditions such as severe combined immunodeficiency,variable immunodeficiency syndrome, and the like.

In some embodiments, a subject having an NK cell-responsive disorder isin need of treatment for bacteremia. Bacteremia is the condition ofbacterial infection in the bloodstream. Septic shock includes seriouslocalized or bacteremic infection accompanied by systemic inflammation,in other words sepsis with hypoperfusion and hypotension refractory tofluid therapy. Sepsis, or systemic inflammatory response syndrome,includes various severe conditions such as infections, pancreatitis,burns, trauma) that can cause acute inflammation. Septic shock istypically related to infections by gram-negative organisms,staphylococci, or meningococci. Septic shock can be characterized byacute circulatory failure, typically with hypotension, and multiorganfailure.

Transient bacteremia can be caused by surgical or trauma wounds.Gram-negative bacteremia can be intermittent and opportunistic; althoughit may have no effect on a healthy person, it may be seriously importantin immunocompromised patients with debilitating underlying diseases,after chemotherapy, and in settings of malnutrition. The infection cantypically be in the lungs, in the genitouritory (GU) or gastrointestinal(GI) tract, or in soft tissues, e.g., skin in patients with decubitusulcer, oral ulcers in patients at risk, and patients with valvular heartdisease, prosthetic heart valves, or other implanted prostheses.

Typically, gram-negative bacteremia can manifest in chronically ill andimmunocompromised patients. Also in such patients, bloodstreaminfections can be caused by aerobic bacilli, anaerobes, and fungi.Bacteroides can lead to abdominal and pelvic infective complications,especially in females. Transient or sustained bacteremia can typicallyresult in metastatic infection of the meninges or serous cavities, suchas the pericardium or larger joints. Enterococcus, staphylococcus, orfungus can lead to endocarditis, but is less common with gram-negativebacteremia. Staphylococcal bacteremia can be typical of IV drug users,and can be a typical cause of gram-positive bacterial endocarditis.

The incidence of systemic fungal infections has undergone a significantincrease, particularly in humans, due in part to increases in the numberof subjects with compromised immune systems, for example, the elderly,AIDS patients, patients undergoing chemotherapy, burn patients, patientswith diabetic ketoacidosis, and transplant patients on immunosuppressivedrugs. A study found that about 40% of deaths from infections acquiredduring hospitalization were due to mycoses; see Sternberg et. al,Science, Vol. 266, (1994), pp. 1632-1634, the entire teachings of whichare incorporated herein by reference.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a fungal infection, such as apathogenic dermatophyte, a pathogenic filamentous fungus, and/or apathogenic non-filamentous fungus, e.g., a yeast, or the like.Pathogenic dermatophytes can include, e.g., species of the generaTrichophyton, Tinea, Microsporum, Epidermophyton, or the like.Pathogenic filamentous fungus can include, e.g., species of genera suchas Aspergillus, Histoplasma, Cryptococcus, Microsporum, or the like.Pathogenic non-filamentous fungus, e.g., yeasts, can include, forexample, species of the genera Candida, Malassezia, Trichosporon,Rhodotorula, Torulopsis, Blastomyces, Paracoccidioides, Coccidioides, orthe like. In various embodiments, the subject can be treated for afungal infection from a species of the genera Aspergillus orTrichophyton. Species of Trichophyton can include, for example,Trichophyton mentagrophytes, Trichophyton rubrum, Trichophytonschoenleinii, Trichophyton tonsurans, Trichophyton verrucosum, andTrichophyton violaceum. Species of Aspergillus can include, for example,Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger,Aspergillus amstelodami, Aspergillus candidus, Aspergillus carneus,Aspergillus nidulans, A. oryzae, Aspergillus restrictus, Aspergillussydowi, Aspergillus terreus, Aspergillus ustus, Aspergillus versicolor,Aspergillus caesiellus, Aspergillus clavatus, Aspergillus avenaceus, andAspergillus deflectus. In some embodiments, the subject can be treatedfor a fungal infection from a pathogenic dermatophyte, e.g.,Trichophyton (e.g., Trichophyton rubrum), Tinea, Microsporum, orEpidermophyton; or Cryptococcus (e.g., Cryptococcus neoformans) Candida(e.g., Candida albicans), Paracoccidioides (e.g., Paracoccidioidesbrasiliensis), or Coccidioides (e.g., Coccidioides immitis). Inparticular embodiments, the subject can be treated for a fungalinfection from Trichophyton rubrum, Cryptococcus neoformans, Candidaalbicans, Paracoccidioides brasiliensis, or Coccidioides immitis.

Thus, in various embodiments, a subject can have an infection caused bya fungus selected from the genera Trichophyton, Tinea, Microsporum,Epidermophyton, Aspergillus, Histoplasma, Cryptococcus, Microsporum,Candida, Malassezia, Trichosporon, Rhodotorula, Torulopsis, Blastomyces,Paracoccidioides, and Coccidioides. In some embodiments, the subject canhave an infection caused by a fungus selected from the generaTrichophyton, Tinea, Microsporum, Epidermophyton; Cryptococcus, Candida,Paracoccidioides, and Coccidioides. In certain embodiments, the subjectcan have an infection caused by a fungus selected from Trichophytonrubrum, Cryptococcus neoformans, Candida albicans, Paracoccidioidesbrasiliensis, and Coccidioides immitis.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a bacterial infection caused,for example, by a bacteria of a genus selected from Allochromatium,Acinetobacter, Bacillus, Campylobacter, Chlamydia, Chlamydophila,Clostridium, Citrobacter, Escherichia, Enterobacter, Enterococcus,Francisella, Haemophilus, Helicobacter, Klebsiella, Listeria, Moraxella,Mycobacterium, Micrococcus, Neisseria, Proteus, Pseudomonas, Salmonella,Serratia, Shigella, Stenotrophomonas, Staphyloccocus, Streptococcus,Synechococcus, Vibrio, and Yersina; or anerobic bacterial genera such asPeptostreptococci, Porphyromonas, Actinomyces, Clostridium, Bacteroides,Prevotella, Anaerobiospirillum, Fusobacterium, and Bilophila. In someembodiments, the subject having an NK cell-responsive disorder can be inneed of treatment for a bacterial infection from Allochromatium vinosum,Acinetobacter baumanii, Bacillus anthracis, Campylobacter jejuni,Chlamydia trachomatis, Chlamydia pneumoniae, Clostridium spp.,Citrobacter spp., Escherichia coli, Enterobacter spp., Enterococcusfaecalis., Enterococcus faecium, Francisella tularensis, Haemophilusinfluenzae, Helicobacter pylori, Klebsiella spp., Listeriamonocytogenes, Moraxella catarrhalis, Mycobacterium tuberculosis,Neisseria meningitidis, Neisseria gonorrhoeae, Proteus mirabilis,Proteus vulgaris, Pseudomonas aeruginosa, Salmonella spp., Serratiaspp., Shigella spp., Stenotrophomonas maltophilia, Staphyloccocusaureus, Staphyloccocus epidermidis, Streptococcus pneumoniae,Streptococcus pyogenes, Streptococcus agalactiae, Yersina pestis, andYersina enterocolitica, or the like; or Peptostreptococciasaccharolyticus, Peptostreptococci magnus, Peptostreptococci micros,Peptostreptococci prevotii, Porphyromonas asaccharolytica, Porphyromonascanoris, Porphyromonas gingivalis, Porphyromonas macaccae, Actinomycesisraelii, Actinomyces odontolyticus, Clostridium innocuum, Clostridiumclostridioforme, Clostridium difficile, Bacteroides tectum, Bacteroidesureolyticus, Bacteroides gracilis (Campylobacter gracilis), Prevotellaintermedia, Prevotella heparinolytica, Prevotella oris-buccae,Prevotella bivia, Prevotella melaminogenica, Fusobacterium naviforme,Fusobacterium necrophorum, Fusobacterium varium, Fusobacterium ulcerans,Fusobacterium russii, Bilophila wadsworthia, Haemophilus ducreyi;Calymmatobacterium granulomatis, or the like.

It is believed that compounds of the invention can be particularlyuseful for treating a subject with an intracellular infection. It isgenerally believed in the art that NK cells are particularly effectiveagainst intracellular infections. Intracellular infections are thosewherein a portion of the infecting pathogen resides within cells of thesubject.

For example, intracellular infections can be caused by one or morebacteria selected from: Ehrlichia (e.g., obligate, intracellularbacteria that can appear as small cytoplasmic inclusions in lymphocytesand neutrophils such as Ehrlichia sennetsu, Ehrlichia canis, Ehrlichiachaffeensis, Ehrlichia phagocytophilia, or the like); Listeria (e.g.,Listeria monocytogenes); Legionella (e.g., Legionella pneumophila);Rickettsiae (e.g., Rickettsiae prowazekii, Rickettsiae typhi(Rickettsiae mooseri), Rickettsiae rickettsii, Rickettsiaetsutsugamushi, Rickettsiae sibirica; Rickettsiae australis; Rickettsiaeconorii; Rickettsiae akari; Rickettsiae burnetii); Chlamydia (e.g.,Chlamydia psittaci; Chlamydia pneumoniae; Chlamydia trachomatis, or thelike); Mycobacterium (Mycobacterium tuberculosis; Mycobacterium marinum;Mycobacterium Avium Complex; Mycobacterium bovis; Mycobacteriumscrofulaceum; Mycobacterium ulcerans; Mycobacterium leprae (Leprosy,Hansen's Bacillus)); Brucella (e.g., Brucella melitensis; Brucellaabortus; Brucella suis; Brucella canis); genus Coxiella (e.g., Coxiellaburnetii); or the like. Thus, in some embodiments, the subject can havean intracellular bacterial infection caused by a bacterium selected fromthe genera Ehrlichia; Listeria; Legionella; Rickettsiae, Chlamydia;Mycobacterium; Brucella; and Coxiella.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a bacterial infection from oneor more upper respiratory tract bacteria. Examples of upper respiratorytract bacteria include those belonging genera such as Legionella,Pseudomonas, and the like. In some embodiments, the bacteria can bePseudomonas aeruginosa. In particular embodiments, the bacteria can beLegionella pneumophila (e.g., including serogroups 1, 2, 3, 4, 5, 6, 7,8, and the like), Legionella dumoffli, Legionella longbeacheae,Legionella micdadei, Legionella oakridgensis, Legionella feelei,Legionella anisa, Legionella sainthelensi, Legionella bozemanii,Legionella gormanii, Legionella wadsworthii, Legionella jordanis, orLegionella gormanii.

In some embodiments, the subject having an NK cell-responsive disordercan be in need of treatment for a bacterial infection from one thatcauses acute bacterial exacerbation of chronic bronchitis (ABECB) in thesubject. Typically, ABECB can be caused by Streptococcus pneumoniae,Haemophilus influenzae, Haemophilus parainfluenzae, or Moraxellacatarrhalis.

In some embodiments, the subject having an NK cell-responsive disordercan be in need of treatment for a bacterial infection from one thatcauses acute community acquired pneumonia (CAP) in the subject.Typically, CAP can be caused by Streptococcus pneumoniae, Haemophilusinfluenzae, Moraxella catarrhalis, Mycoplasma pneumoniae, Chlamydiapneumoniae, or Klebsiella pneumoniae. In a particular embodiment, theCAP can be caused by drug resistant bacteria, e.g., a multi-drugresistant strain of Streptococcus pneumoniae.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a bacterial infection fromStreptococcus pneumoniae, Haemophilus influenzae, Haemophilusparainfluenzae, Moraxella catarrhalis, Mycoplasma pneumoniae, Chlamydiapneumoniae, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcuspyogenes, Acinetobacter lwoffi, Klebsiella oxytoca, Legionellapneumophila, or Proteus vulgaris.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a bacterial infection frommaxillary sinus pathogenic bacteria. As used herein, maxillary sinuspathogenic bacteria is a bacterial strain isolated from acute or chronicmaxillary sinusitis, or, for example, a maxillary sinus isolate ofStaphylococcus aureus, Streptococcus pneumoniae, Haemophilus spp.,Moraxella catarrhalis, an anaerobic strain of non-fermentative Gramnegative bacilli, Neisseria meningitides or β-haemolytic Streptococcus.In various embodiments, maxillary sinus pathogenic bacteria can includea bacterial strain isolated from acute or chronic maxillary sinusitis; amaxillary sinus isolate of Staphylococcus aureus, Streptococcuspneumoniae, Haemophilus spp., Moraxella catarrhalis, an anaerobic strainof non-fermentative Gram negative bacilli, Neisseria meningitidis,β-haemolytic. Streptococcus, Haemophilus influenzae, anEnterobacteriaceae, a non-fermentative Gram negative bacilli,Streptococcus pneumoniae, Streptococcus pyogenes, amethicillin-resistant Staphylococcus spp., Legionella pneumophila,Mycoplasma spp. and Chlamydia spp., Haemophilus influenzae, Haemophilusparainfluenzae, Peptostreptococcus, Bacteroides spp., and Bacteroidesurealyticus.

In various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for a bacterial infection thatcauses a urinary tract infection (UTI) in the subject. Examples of UTIsinclude urethritis, cystitis, prostatitis, pyelonephritis (acute,chronic, and xanthogranulomatous), and hematogenous UTI (e.g., frombacteremia with virulent bacilli such as Salmonella, Staphylococcusaureus, and the like). Typically, UTIs can be caused by gram-negativeaerobic bacteria, e.g., Escherichia (e.g., Escherichia coli),Klebsiella, Proteus, Enterobacter, Pseudomonas, and Serratia;gram-negative anaerobic bacteria; gram-positive bacteria, e.g.,Enterococci (e.g., Enterococcus faecalis) and Staphylococcus sp (e.g.,Staphylococcus saprophyticus, Staphylococcus aureus, and the like);Mycobacterium tuberculosis; and sexually transmitted bacterialinfections (e.g., Chlamydia trachomatis, Neisseria gonorrhoeae, and thelike).

In certain embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for infections from microorganismsthat cause sexually transmitted diseases, for example, Treponemapallidum; Trichomonas vaginalis; Candidia (Candida albicans); Neisseriagonorrhoeae; Chlamydia trachomatis; Mycoplasma genitalium, Ureaplasmaurealyticum; Haemophilus ducreyi; Calymmatobacterium granulomatis(formerly Donovania granulomatis); herpes simplex viruses (HSV-1 orHSV-2); human papillomavirus [HPV]; human immunodeficiency virus (HIV);various bacterial (Shigella, Campylobacter, or Salmonella), viral(hepatitis A), or parasitic (Giardia or amoeba, e.g., Entamoeba dispar(previously Entamoeba histolytica); or the like.

Thus, in various embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for an infection resulting in upperrespiratory tract bacterial infection, acute bacterial exacerbation ofchronic bronchitis; acute community acquired pneumonia, maxillary sinuspathogenic bacteria; a urinary tract infection; or a sexuallytransmitted infection.

It is believed that the methods can be particularly effective fortreating a subject with a viral infection. Thus, in various embodiments,a subject having an NK cell-responsive disorder can be in need oftreatment for infection from viruses such as Picornaviruses (e.g., PolioVirus, rhinoviruses and certain echoviruses and coxsackieviruses);Parvoviridae (Human Parvovirus B19); Hepatitis, e.g, Hepadnavirus(Hepatitis B); Papovavirus (JC Virus); Adenovirus (Human Adenovirus);Herpesvirus (e.g., Cytomegalovirus, Epstein Barr Virus (Mononucleosis),Mononucleosis-Like Syndrome, Roseola Infantum, Varicella Zoster Virus(Chicken Pox), Herpes Zoster (Shingles), Herpes Simplex Virus (OralHerpes, Genital Herpes)), Poxvirus (Smallpox); Calicivirus (NorwalkVirus), Arbovirus (e.g., Togavirus (Rubella virus, Dengue virus),Flavivirus (Yellow Fever virus), Bunyavirus (California EncephalitisVirus), Reovirus (Rotavirus)); Coronavirus (Coronavirus); Retrovirus(Human Immunodeficiency Virus 1, Human Immunodeficiency Virus 2);Rhabdovirus (Rabies Virus), Filovirus (Marburg Virus, Ebola virus, otherhemorrhagic viral diseases); Paramyxovirus (Measles Virus, Mumps Virus);Orthomyxovirus (Influenza Virus); Arenavirus (Lassa Fever); human T-cellLymphotrophic virus type I and II (HTLV-I, HTLV II); humanpapillomavirus [HPV]; or the like. Thus, in various embodiments, thesubject can have an infection caused by a virus selected fromPicornavirus; Parvoviridae; Hepatitis virus; Papovavirus; Adenovirus;Herpesvirus, Poxvirus; Calicivirus; Arbovirus; Coronavirus; aRetrovirus; Rhabdovirus; Paramyxovirus; Orthomyxovirus; Arenavirus;human T-cell Lymphotrophic virus; human papillomavirus; and humanimmunodeficiency virus.

In some embodiments, a subject having an NK cell-responsive disorder canbe in need of treatment for an infection from a virus or an infectionthereof such as human immunodeficiency virus-1, human immunodeficiencyvirus-2, Cytomegalovirus, Epstein Barr Virus, Mononucleosis-LikeSyndrome, Roseola Infantum, Varicella Zoster Virus, Herpes Zoster,Herpes Simplex Virus, or hepatitis.

It is believed that the methods can be particularly effective fortreating a subject with a parasitic infection. Thus, in variousembodiments, a subject having an NK cell-responsive disorder can be inneed of treatment for an infection from Plasmodia (e.g., Plasmodiafalciparum, Plasmodia vivax, Plasmodia ovale, and Plasmodia malariae,typically transmitted by anopheline mosquitoes); Leishmania (transmittedby sandflies and caused by obligate intracellular protozoa, e.g.,Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmaniamexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmaniatropica; Leishmania major; Leishmania aethiopica; and the subgenusViannia, Leishmania Viannia braziliensis, Leishmania Viannia guyanensis,Leishmania Viannia panamensis, and Leishmania Viannia peruviana);Trypanosoma (e.g., sleeping sickness caused by Trypanosoma bruceigambiense, and Trypanosoma brucei rhodesiense); amoebas of the generaNaegleria or Acanthamoeba; pathogens such as genus Entamoeba (Entamoebahistolytica and Entamoeba dispar); Giardia lamblia; Cryptosporidium;Isospora; Cyclospora; Microsporidia; Ascaris lumbricoides; infectionwith blood flukes of the genus Schistosoma (e.g.; S. haematobium; S.mansoni; S. japonicum; S. mekongi; S. intercalatum); Toxoplasmosis(e.g., Toxoplasma gondii); Treponema pallidum; Trichomonas vaginalis; orthe like.

In some embodiments, the subject having an NK cell-responsive disordercan have an infection caused by a protozoa selected from Toxoplasmagondii, Trypanosoma brucei gambiense, Trypanosoma brucei rhodesiense,Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmaniamexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmaniatropica; Leishmania major; Leishmania aethiopica; and the subgenusViannia, Leishmania Viannia braziliensis, Leishmania Viannia guyanensis,Leishmania Viannia panamensis, Leishmania Viannia peruviana, Plasmodiafalciparum, Plasmodia vivax, Plasmodia ovale, and Plasmodia malariae.

In the last century, antibiotics were developed that led to significantreductions in mortality. Unfortunately, widespread use has led to therise of antibiotic resistant bacteria, e.g., methicillin resistantStaphyloccocus aureus (MRSA), vancomycin resistant enterococci (VRE),and penicillin-resistant Streptococcus pneumoniae (PRSP). Some bacteriaare resistant to a range of antibiotics, e.g., strains of Mycobacteriumtuberculosis resist isoniazid, rifampin, ethambutol, streptomycin,ethionamide, kanamycin, and rifabutin. In addition to resistance, globaltravel has spread relatively unknown bacteria from isolated areas to newpopulations. Furthermore, there is the threat of bacteria as biologicalweapons. These bacteria may not be easily treated with existingantibiotics.

It is believed that the compounds of the invention can be particularlyeffective for treating a subject for drug-resistant pathogens, forexample, drug resistant bacteria, or pathogens for which no drugs areavailable, e.g., many viruses. Without wishing to be bound by theory, itis believed that because the compounds of the invention can act byincreasing NK cell activity, and thus the NK cells can kill infectivemicroorganisms or infected cells separately from any direct action ofthe compounds on the pathogen or infected cells. Thus, it is believedthat the compounds of the invention can have at least one mode of actionthat is separate from typical anti-infective drugs such as antibioticswhich can typically act directly on the bacteria themselves.

Drug resistant pathogens can be resistant to at least one and typicallymultiple agents, for example, drug resistant bacteria can be resistantto one antibiotic, or typically at least two antibiotics such aspenicillin, Methicillin, second generation cephalosporins (e.g.,cefuroxime, and the like), macrolides, tetracyclines,trimethoprim/methoxazole, vancomycin, or the like. For example, in someembodiments, a subject can be treated for bacteria selected from astrain of multiple drug resistant Streptococcus pneumoniae (MDRSP,previously known as penicillin resistant Streptococcus pneumoniae,PRSP), vancomycin resistant Enterococcus, methicillin resistantStaphylococcus Aureus, penicillin resistant Pneumococcus, antibioticresistant Salmonella, resistant and multi-resistant Neisseria Gonorrhea(e.g., resistant to one, two or more of tetracycline, penicillin,fluoroquinolones, cephalosporins, ceftriaxone (Rocephin), Cefixime(Suprax), Azithromycin, or the like), and resistant and multi-resistantTuberculosis (e.g., resistant to one, two or more of Isoniazid,Rifampin, Ethambutol, Pyrazinamide, Aminoglycoside, Capreomycin,Ciprofloxacin, Ofloxacin, gemifloxacin, Cycloserine, Ethionamide,para-aminosalicylic acid or the like).

In some embodiments, NK cell activity can be increased in subjects thathave an immunodeficiency. In various embodiments, this can be due todecreased or deficient NK cell activity. In some embodiments, theimmunodeficiency can be any known immunodeficiency, even those that donot directly impact NK cells. Without wishing to be bound by theory, itis believed that boosting NK cell activity can augment immune functionin many immunodeficiency conditions to “make-up” at least in part, foraspects of immunodeficiency separate from those aspects directlyconcerned with NK cell activity.

In various embodiments, immunodeficiency disorders can include disorderswith increased susceptibility to infection, for example, one or moredisorders selected from: circulatory and systemic disorders (sickle celldisease, diabetes mellitus, nephrosis, varicose veins, congenitalcardiac defects); obstructive disorders (ureteral or urethral stenosis,bronchial asthma, bronchiectasis, allergic rhinitis, blocked Eustachiantubes); integumentary defects (eczema, burns, skull fractures, midlinesinus tracts, ciliary abnormalities); primary immunodeficiencies(X-linked agammaglobulinemia, DiGeorge anomaly, chronic granulomatousdisease, C3 deficiency); secondary immunodeficiencies (malnutrition,prematurity, lymphoma, splenectomy, uremia, immunosuppressive therapy,protein-losing enteropathy, chronic viral diseases); unusualmicrobiologic factors (antibiotic overgrowth, chronic infections withresistant organism, continuous reinfection (contaminated water supply,infectious contact, contaminated inhalation therapy equipment)); foreignbodies, trauma (ventricular shunts, central venous catheter, artificialheart valves, urinary catheter, aspirated foreign bodies) allogeneictransplant, graft-versus-host disease, uterine dysfunction (e.g.,endometriosis), or the like.

In various embodiments, immunodeficiency disorders can include forexample, transient hypogammaglobulinemia of infancy, selective IgAdeficiency, X-linked agammaglobulinemian (Bruton's Agammaglobulinemia;Congenital Agammaglobulinemia), common variable immunodeficiency(Acquired Agammaglobulinemia), hyper-IgM immunodeficiency, IgG subclassdeficiency, chronic mucocutaneous Candidiasis, combinedimmunodeficiency, Wiskott-Aldrich syndrome, ataxia-telangiectasia,X-linked lymphoproliferative syndrome, hyper-IgE syndrome (Job-BuckleySyndrome), chronic granulotomatous disease, leukocyte adhesiondeficiency (MAC-1/LFA-1/CR3 deficiency), or the like.

In various embodiments, immunodeficiency disorders can include primaryimmunodeficiency disorders for example: B-cell (antibody) deficiencies(X-linked agammaglobulinemia; Ig deficiency with hyper-IgM (XL); IgAdeficiency); IgG subclass deficiencies, Antibody deficiency with normalor elevated Igs, Immunodeficiency with theymoma, Common variableimmunodeficiency, Transient hypogammaglobulinemia of infancy); T-cell(cellular) deficiencies (Predominant T-cell deficiency: DiGeorgeanomaly, Chronic mucocutaneous candidiasis, Combined immunodeficiencywith Igs (Nezelof syndrome), Nucleoside phosphorylase deficiency (AR),Natural killer cell deficiency, Idiopathic CD4 lymphocytopenia, CombinedT- and B-cell deficiencies: Severe combined immunodeficiency (AR or XL),Adenosine deaminase deficiency (AR), Reticular dysgenesis, Barelymphocyte syndrome, Ataxia-telangiectasia (AR), Wiskott-Aldrichsyndrome (XL), Short-limbed dwarfism, XL lymphoproliferative syndrome);Phagocytic disorders (Defects of cell movement: HyperimmunoglobulinemiaE syndrome, Leukocyte adhesion defect type 1 (AR), Defects ofmicrobicidal activity: Chronic granulomatous disease (XL or AR),Neutrophil G6PD deficiency, Myeloperoxidase deficiency (AR),Chediak-Higashi syndrome (AR)); Complement disorders (Defects ofcomplement components: C1q deficiency, Defects of control proteins: C1inhibitor deficiency (D1), Factor I (C3b inactivator) deficiency (ACD),Factor H deficiency (ACD), Factor D deficiency (ACD), Properdindeficiency (XL)); or the like

In various embodiments, immunodeficiency disorders can include secondaryimmunodeficiency disorders, for example, one or more conditions selectedfrom: Premature and newborn infants (Physiologic immunodeficiency due toimmaturity of immune system); Hereditary and metabolic diseases(Chromosome abnormalities (e.g., Down syndrome), Uremia, Diabetes (i.e.,complications from diabetes such as gangrene associated with peripheralcirculatory and nerve dysfunction), Malnutrition, Vitamin and mineraldeficiencies, Protein-losing enteropathies, Nephrotic syndrome, Myotonicdystrophy, Sickle cell disease); Immunosuppressive agents (Radiation,Immunosuppressive drugs, Corticosteroids, Anti-lymphocyte oranti-thymocyte globulin, Anti-T-cell monoclonal antibodies); Infectiousdiseases (Congenital rubella, Viral exanthems (e.g., measles,varicella), HIV infection, Cytomegalovirus infection, Infectiousmononucleosis, Acute bacterial disease, Severe mycobacterial or fungaldisease); Infiltrative and hematologic diseases (Histiocytosis,Sarcoidosis, Hodgkin's disease and lymphoma, Leukemia, Myeloma,Agranulocytosis and aplastic anemia); Surgery and trauma (Burns,Splenectomy, Anesthesia, wounds); and Miscellaneous (SLE, Chronic activehepatitis, Alcoholic cirrhosis, Aging, Anticonvulsive drugs,Graft-vs.-host disease); or the like.

In certain embodiments, the subject having an NK cell-responsivedisorder can be in need of treatment for burns or wounds. Typically,such a wound or burn is a severe injury that places a significant burdenon the subject's immune defenses. For example, in some embodiments, thesubject is treated for a second or third degree burn covering at leastabout 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 75%, or more of the surfacearea of the subject's body. Also, in some embodiments, the subject istreated for a wound or wounds, e.g., an open wound of at least about 1cm², 2 cm², 5 cm², 10 cm², 20 cm², 50 cm² or larger, or 1%, 2%, 3%, 4%,5%, 10%, 15%, or more of the surface area of the subject's body; or oneor more incisions penetrating the skin totaling at least 1 cm, 2 cm, 3cm, 4 cm, 5 cm, 7 cm, 10 cm, 20 cm, 25 cm, 50 cm in length; anamputation; and the like.

In various embodiments, the subject having an NK cell-responsivedisorder can have an infection caused by antibiotic resistant bacteria.In some embodiments, the subject can have an infection caused by abacterium selected from multiple drug resistant Streptococcuspneumoniae, vancomycin resistant Enterococcus, methicillin resistantStaphylococcus Aureus, penicillin resistant Pneumococcus, antibioticresistant Salmonella, resistant/multi-resistant Neisseria Gonorrhea, andresistant/multi-resistant Tuberculosis. In some embodiments, the subjectcan have a bacterial infection resistant to at least one antibioticselected from penicillin, Methicillin, second generation cephalosporins,macrolides, tetracyclines, trimethoprim/methoxazole, vancomycin,tetracycline, fluoroquinolones, ceftriaxone, Cefixime, Azithromycin,Isoniazid, Rifampin, Ethambutol, Pyrazinamide, Aminoglycoside,Capreomycin, Ciprofloxacin, Ofloxacin, gemifloxacin, Cycloserine,Ethionamide, and para-aminosalicylic acid.

Thus, various embodiments, the subject having an NK cell responsivedisorder can have an immunodeficiency disorder. In some embodiments, thesubject can have a primary immunodeficiency disorder. In someembodiments, the subject can have a secondary immunodeficiency disorder.

In some embodiments, immunodeficiency disorders can include uremia,diabetes (infective complications thereof, malnutrition, vitamin andmineral deficiencies, protein-losing enteropathies, nephrotic syndrome,myotonic dystrophy, sickle cell disease; or the like.

In some embodiments, immunodeficiency disorders can be cause or bepartially caused by immunosuppressive agents, e.g., radiation,immunosuppressive drugs, corticosteroids, anti-lymphocyte oranti-thymocyte globulin, anti-T-cell monoclonal antibodies; or the like.

In some embodiments, immunodeficiency disorders can caused or partiallycaused by surgery and trauma, e.g., burns, splenectomy, anesthesia,wounds, implanted medical devices; or the like.

In some embodiments, immunodeficiency disorders can include chronicfatigue syndrome (chronic fatigue immune dysfunction syndrome);Epstein-Barr virus infection, post viral fatigue syndrome,post-transplantation syndrome (host-graft disease), exposure to nitricoxide synthase inhibitors, aging, severe combined immunodeficiency,variable immunodeficiency syndrome, and the like.

Increasing NK cell activity would also be beneficial for treatingsubjects with disorders including, but not limited to aneurodegenerative disorder. As used herein, a neurodegenerative disorderinvolves degradation of neurons such as cerebral, spinal, and peripheralneurons (e.g., at neuromuscular junctions), more typically degradationof cerebral and spinal neurons. Neurodegenerative disorders can includeAlzheimer's disease; Huntington's disease; Parkinson's disease;spinal/bulbar muscular atrophy (e.g., Kennedy's disease),spinocerebellar ataxic disorders, and other neuromuscular atrophies;familial amyotrophic lateral sclerosis; ischemia; seizure; hypothermia;hyperthermia; burn trauma; atherosclerosis; radiation exposure;glaucoma; toxin exposure; mechanical injury; inflammation; epilepticseizure, injury-induced seizure, chemically-induced seizure, or otherdiseases associated with superoxide dismutase (SOD) mutations; and thelike. Neurodegenerative disorders can also include degradation ofneurons caused by ischemia, seizure, thermal stress, radiation, toxinexposure, infection, injury, and the like. Ischemia can damage tissuethrough multiple routes, including oxygen depletion, glucose depletion,oxidative stress upon reperfusion, and/or glutamate toxicity, and thelike. Ischemia can result from an endogenous condition (e.g., stroke,heart attack, and the like), from accidental mechanical injury, fromsurgical injury (e.g., reperfusion stress on transplanted organs), andthe like. Alternatively, tissues that can be damaged by ischemia includeneurons, cardiac muscle, liver tissue, skeletal muscle, kidney tissue,pulmonary tissue, pancreatic tissue, and the like.

Other disorders in which increasing NK cell activity would be beneficialinclude disorders due to thermal stress, (thermal stress includeshyperthermia (e.g., from fever, heat stroke, burns, and the like) andhypothermia); radiation damage, e.g., due to visible light, ultravioletlight, microwaves, cosmic rays, alpha radiation, beta radiation, gammaradiation, X-rays, and the like, (for example, the damage could beradiation damage to non-cancerous tissue in a subject treated for cancerby radiation therapy); mechanical injury, e.g., trauma from surgery,accidents, certain disease conditions (e.g., pressure damage inglaucoma) and the like; and exposure to a toxin. e.g., exposure to aneurotoxin selected from methamphetamine; antiretroviral HIVtherapeutics (e.g., nucleoside reverse transcriptase inhibitors; heavymetals (e.g., mercury, lead, arsenic, cadmium, compounds thereof, andthe like), amino acid analogs, chemical oxidants, ethanol, glutamate,metabolic inhibitors, antibiotics, and the like.

Another embodiment of the present invention is a method of treating asubject with a cancer. Optionally, the method of the invention can beused for a multi-drug resistant cancer as described below. The methodcomprises the step of administering an effective amount of a compound offormula (I) through (VII) and Table 1, or a tautomer, pharmaceuticallyacceptable salt, solvate, clathrate, or a prodrug thereof. Preferably,one or more additional anti-cancer drugs are co-administered with acompound of the invention. Examples of anti-cancer drugs are describedbelow. Preferably, the co-administered anti-cancer drug is an agent thatstabilizes microtubules, such as Taxol® or a taxanes derivative.

As noted above, one embodiment of the present invention is directed totreating subjects with a cancer. “Treating a subject with a cancer”includes achieving, partially or substantially, one or more of thefollowing: arresting the growth or spread of a cancer, reducing theextent of a cancer (e.g., reducing size of a tumor or reducing thenumber of affected sites), inhibiting the growth rate of a cancer, andameliorating or improving a clinical symptom or indicator associatedwith a cancer (such as tissue or serum components).

In another embodiment, a compound of the invention can be administeredas adjuvant therapy to prevent the reoccurrence of cancer. For example,stage II and stage III melanoma are typically treated with surgery toremove the melanoma followed by chemotherapeutic treatment to preventthe reoccurrence of cancer. In one embodiment, one or more additionalanti-cancer drugs are co-administered with a compound of the inventionas adjuvant therapy. Examples of anti-cancer drugs are described below.In one embodiment, the co-administered anti-cancer drug is an agent thatstabilizes microtubules, such as Taxol® or a taxanes derivative. Inanother embodiment, the co-administered anti-cancer drug is animmunotherapeutic anticancer agent.

Cancers that can be treated or prevented by the methods of the presentinvention include, but are not limited to human sarcomas and carcinomas,e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenicsarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor,leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, colorectal cancer,anal carcinoma, esophageal cancer, gastric cancer, hepatocellularcancer, bladder cancer, endometrial cancer, pancreatic cancer, breastcancer, ovarian cancer, prostate cancer, stomach cancer, atrial myxomas,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, thyroid and parathyroidneoplasms, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testiculartumor, lung carcinoma, small cell lung carcinoma, non-small-cell lungcancer, bladder carcinoma, epithelial carcinoma, glioma, pituitaryneoplasms, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma,pinealoma, hemangioblastoma, acoustic neuroma, schwannomas,oligodendroglioma, meningioma, spinal cord tumors, melanoma,neuroblastoma, pheochromocytoma, Types 1-3 endocrine neoplasia,retinoblastoma; leukemias, e.g., acute lymphocytic leukemia and acutemyelocytic leukemia (myeloblastic, promyelocytic, myelomonocytic,monocytic and erythroleukemia); chronic leukemia (chronic myelocytic(granulocytic) leukemia and chronic lymphocytic leukemia); andpolycythemia vera, lymphoma (Hodgkin's disease and non-Hodgkin'sdisease), multiple myeloma, Waldenstrobm's macroglobulinemia, and heavychain disease.

Other examples of leukemias include acute and/or chronic leukemias,e.g., lymphocytic leukemia (e.g., as exemplified by the p388 (murine)cell line), large granular lymphocytic leukemia, and lymphoblasticleukemia; T-cell leukemias, e.g., T-cell leukemia (e.g., as exemplifiedby the CEM, Jurkat, and HSB-2 (acute), YAC-1(murine) cell lines),T-lymphocytic leukemia, and T-lymphoblastic leukemia; B cell leukemia(e.g., as exemplified by the SB (acute) cell line), and B-lymphocyticleukemia; mixed cell leukemias, e.g., B and T cell leukemia and B and Tlymphocytic leukemia; myeloid leukemias, e.g., granulocytic leukemia,myelocytic leukemia (e.g., as exemplified by the HL-60 (promyelocyte)cell line), and myelogenous leukemia (e.g., as exemplified by theK562(chronic)cell line); neutrophilic leukemia; eosinophilic leukemia;monocytic leukemia (e.g., as exemplified by the THP-1 (acute) cellline); myelomonocytic leukemia; Naegeli-type myeloid leukemia; andnonlymphocytic leukemia. Other examples of leukemias are described inChapter 60 of The Chemotherapy Sourcebook, Michael C. Perry Ed.,Williams & Williams (1992) and Section 36 of Holland Frie CancerMedicine 5th Ed., Bast et al. Eds., B. C. Decker Inc. (2000). The entireteachings of the preceding references are incorporated herein byreference.

Additional cancers that can be treated or prevented by the methods ofthe present invention include, but are not limited to oral cavity &pharynx cancers, including tongue, mouth, pharynx, and other oral cavitycancers; digestive system cancers, including esophagus, small intestine,rectum, anus, anal canal, anorectum, liver & intrahepatic bile duct,gallbladder & other biliary, pancreas and other digestive organs;respiratory system cancers, including larynx and bronchus; bone & jointcancers; soft tissue (including heart) cancers; genital system cancers,including uterine cervix, uterine corpus, ovary, vulva, vagina & othergenital, female, testis, penis & other genital, male; urinary systemcancers, including kidney & renal pelvis, and ureter & other urinaryorgans; eye & orbit cancers; leukemia, including acute myeloid leukemiaand chronic myeloid leukemia.

In one embodiment, the disclosed method is believed to be particularlyeffective in treating a subject with non-solid tumors such as multiplemyeloma. In another embodiment, the disclosed method is believed to beparticularly effective against T-leukemia (e.g., as exemplified byJurkat and CEM cell lines); B-leukemia (e.g., as exemplified by the SBcell line); promyelocytes (e.g., as exemplified by the HL-60 cell line);uterine sarcoma (e.g., as exemplified by the MES-SA cell line);monocytic leukemia (e.g., as exemplified by the THP-1 (acute) cellline); and lymphoma (e.g., as exemplified by the U937 cell line).

In another embodiment, the disclosed method is believed to beparticularly effective in treating a subject with an immunosensitivecancer. Immunosensitive cancers are cancers that respond to treatmentwith immunotherapy. Immunotherapy is described below in more detail.Cancers that respond to immunotherapy include renal cell carcinoma,melanoma (including superficial spreading melanoma, nodular melanoma,acral lentiginous melanoma, lentigo maligna melanoma which is alsocalled Hutchinson's Freckle), multiple myeloma, myeloma, lymphoma,non-small-cell lung cancer, squamous cell carcinoma, basal cellcarcinoma, fibrosarcoma and malignant brain tumors.

In another embodiment, the disclosed method is believed to beparticularly effective in treating a subject with melanoma.

In another embodiment, the disclosed method is believed to beparticularly effective in treating a subject with renal cell carcinoma.

The disclosed method is particularly effective at treating subjectswhose cancer has become “multi-drug resistant”. A cancer which initiallyresponded to an anti-cancer drug becomes resistant to the anti-cancerdrug when the anti-cancer drug is no longer effective in treating thesubject with the cancer. For example, many tumors will initially respondto treatment with an anti-cancer drug by decreasing in size or evengoing into remission, only to develop resistance to the drug. Drugresistant tumors are characterized by a resumption of their growthand/or reappearance after having seemingly gone into remission, despitethe administration of increased dosages of the anti-cancer drug. Cancersthat have developed resistance to two or more anti-cancer drugs are saidto be “multi-drug resistant”. For example, it is common for cancers tobecome resistant to three or more anti-cancer agents, often five or moreanti-cancer agents and at times ten or more anti-cancer agents.

Numerous non-cancer diseases involve excessive or hyperproliferativecell growth, termed hyperplasia. As used herein, the terms“proliferative disorder”, “hyperproliferative disorder,” and “cellproliferation disorder” are used interchangeably to mean a disease ormedical condition involving pathological growth of cells. Such disordersinclude cancer.

Non-cancerous proliferative disorders include smooth muscle cellproliferation, systemic sclerosis, cirrhosis of the liver, adultrespiratory distress syndrome, idiopathic cardiomyopathy, lupuserythematosus, retinopathy, e.g., diabetic retinopathy or otherretinopathies, cardiac hyperplasia, reproductive system associateddisorders such as benign prostatic hyperplasia and ovarian cysts,pulmonary fibrosis, endometriosis, fibromatosis, hematomas,lymphangiomatosis, sarcoidosis, desmoid tumors and the like.

Smooth muscle cell proliferation includes proliferative vasculardisorders, for example, intimal smooth muscle cell hyperplasia,restenosis and vascular occlusion, particularly stenosis followingbiologically- or mechanically-mediated vascular injury, e.g., vascularinjury associated with balloon angioplasty or vascular stenosis.Moreover, intimal smooth muscle cell hyperplasia can include hyperplasiain smooth muscle other than the vasculature, e.g., hyperplasia in bileduct blockage, in bronchial airways of the lung in asthma patients, inthe kidneys of patients with renal interstitial fibrosis, and the like.

Non-cancerous proliferative disorders also include hyperproliferation ofcells in the skin such as psoriasis and its varied clinical forms,Reiter's syndrome, pityriasis rubra pilaris, and hyperproliferativevariants of disorders of keratinization (e.g., actinic keratosis, senilekeratosis), scleroderma, and the like.

An “effective amount” is the quantity of compound in which a beneficialclinical outcome is achieved when the compound is administered to asubject. For example, when a compound of the invention is administeredto a subject with a cancer, a “beneficial clinical outcome” includes areduction in tumor mass, a reduction in metastasis, a reduction in theseverity of the symptoms associated with the cancer and/or an increasein the longevity of the subject compared with the absence of thetreatment. When a compound of the invention is administered to a subjectwith a an Hsp70-responsive disorder or an NK cell-responsive disorder, a“beneficial clinical outcome” includes reduction in the severity ornumber of symptoms associated with the disorder, elimination of aninfection, or increase in the longevity of the subject compared with theabsence of the treatment. The precise amount of compound administered toa subject will depend on the type and severity of the disease orcondition and on the characteristics of the subject, such as generalhealth, age, sex, body weight and tolerance to drugs. It may also dependon the degree, severity and type of cancer. The skilled artisan will beable to determine appropriate dosages depending on these and otherfactors. Effective amounts of the disclosed compounds typically rangebetween about 1 mg/m² per day and about 10 grams/m² per day, andpreferably between 10 mg/m² per day and about 5 grams/m². Whenco-administered with another anti-cancer agent for the treatment ofcancer, an “effective amount” of the second anti-cancer agent willdepend on the type of drug used. Suitable dosages are known for approvedanti-cancer agents and can be adjusted by the skilled artisan accordingto the condition of the subject, the type of cancer being treated andthe compound of the invention being used.

Another embodiment of the present invention is a pharmaceuticalcomposition comprising a compound of the invention, or a tautomer,pharmaceutically acceptable salt, solvate, clathrate, or a prodrugthereof, and a pharmaceutically acceptable carrier or diluent.

Suitable pharmaceutically acceptable carriers may contain inertingredients which do not inhibit the biological activity of thedisclosed disalts. The pharmaceutically acceptable carriers should bebiocompatible, i.e., non-toxic, non-inflammatory, non-immunogenic anddevoid of other undesired reactions upon the administration to asubject. Standard pharmaceutical formulation techniques can be employed,such as those described in Remington's Pharmaceutical Sciences, MackPublishing Company, Easton, Pa. Formulation of the compound to beadministered will vary according to the route of administration selected(e.g., solution, emulsion, capsule). Suitable pharmaceutical carriersfor parenteral administration include, for example, sterile water,physiological saline, bacteriostatic saline (saline containing about0.9% mg/ml benzyl alcohol), phosphate-buffered saline, Hank's solution,Ringer's-lactate and the like. Methods for encapsulating compositions(such as in a coating of hard gelatin or cyclodextrins) are known in theart (Baker, et al., “Controlled Release of Biological Active Agents”,John Wiley and Sons, 1986).

Other anti-proliferative or anticancer therapies may be combined withthe compounds of this invention to treat proliferative diseases andcancer. Other therapies or anticancer agents that may be used incombination with the inventive anticancer agents of the presentinvention include surgery, radiotherapy (including, but not limited to,gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy,proton therapy, brachytherapy, and systemic radioactive isotopes),endocrine therapy, biologic response modifiers (including, but notlimited to, interferons, interleukins, and tumor necrosis factor (TNF)),hyperthermia and cryotherapy, agents to attenuate any adverse effects(e.g., antiemetics), and other approved chemotherapeutic drugs.

The prophylactic or therapeutic agents of the combination therapies ofthe invention can be administered sequentially or concurrently.

As used herein, the terms “hyperthermia”, “hyperthermia therapy,”“thermal therapy,” and “thermotherapy” are used interchangeably to meana treatment where body tissue is exposed to high temperatures (up to113° F.). The term as used herein includes all forms of hyperthermia,including local, regional, and whole-body. Various forms of energy canbe used to deliver heat to the desired area, such as microwave,radiofrequency, lasers, and ultrasound. The treatment temperatures varydepending on the location of the tumor and the approach used.

In local hyperthermia, heat is applied to a small area (e.g. a tumor).The approaches to local hyperthermia vary with tumor location. Externalapproaches are used to treat tumors in or just below the skin. In thismethod, applicators are place near or around the tumor and deliverenergy directly to the tumor. Intraluminal or endocavitary approachesuse probes to deliver energy to tumors within or near body cavities.Interstitial approaches are used to treat tumors deep within the body(e.g. brain tumors), by inserting probes or needles into the tumor underanesthesia.

In regional hyperthermia, heat is applied to large areas of tissue (e.g.body cavity, organ, or limb). Deep tissue approaches are used to treatcancers within the body (e.g. cervical or bladder cancer) by usingexternal applicators. Regional perfusion approaches are used to treatcancers in the limbs or organs (e.g. melanoma, liver, or lung cancer).In this approach some of the blood is removed and heated and then pumpedback into the limb or organ. Anticancer drugs may be given during thisprocess. Continuous hyperthermic peritoneal perfusion (CHPP) is used totreat cancers in the peritoneal cavity (e.g. peritoneal mesothelioma orstomach cancer). In this approach, heated anticancer drugs are pumpedthrough the peritoneal cavity.

Whole-body hyperthermia is used to treat metastatic cancer. In thisapproach, the whole body is heated to 107-108° F. by using varioustechniques such as thermal chambers or hot water blankets.

Hyperthermic conditions are known to induce the synthesis of Hsp70.

The compounds of the invention are administered by any suitable route,including, for example, orally in capsules, suspensions or tablets or byparenteral administration. Parenteral administration can include, forexample, systemic administration, such as by intramuscular, intravenous,subcutaneous, or intraperitoneal injection. The compounds of theinvention can also be administered orally (e.g., dietary), topically, byinhalation (e.g., intrabronchial, intranasal, oral inhalation orintranasal drops), or rectally, depending on the type of cancer to betreated. Oral and parenteral administrations are preferred modes ofadministration.

Many new drugs are now available to be used by oncologists in treatingpatients with cancer. Often, tumors are more responsive to treatmentwhen anti-cancer drugs are administered in combination to the patientthan when the same drugs are administered individually and sequentially.One advantage of this approach is that the anti-cancer agents often actsynergistically because the tumors cells are attacked simultaneouslywith agents having multiple modes of action. Thus, it is often possibleto achieve more rapid reductions in tumor size by administering thesedrugs in combination. Another advantage of combination chemotherapy isthat tumors are more likely to be eradicated completely and are lesslikely to develop resistance to the anti-cancer drugs being used totreat the patient.

Optionally, a compound of the invention, or a tautomer, pharmaceuticallyacceptable salt, solvate, clathrate, or a prodrug thereof, can beco-administered to treat a patient with a proliferative disorder such ascancer, or to prevent the reoccurrence of a proliferative disorder suchas cancer, with other anti-cancer agents such as Adriamycin,Dactinomycin, Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin;acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine;ambomycin; ametantrone acetate; aminoglutethimide; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin;dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicinhydrochloride; droloxifene; droloxifene citrate; dromostanolonepropionate; duazomycin; edatrexate; eflornithine hydrochloride;elsamitrucin; enloplatin; enpromate; epipropidine; epirubicinhydrochloride; erbulozole; esorubicin hydrochloride; estramustine;estramustine phosphate sodium; etanidazole; etoposide; etoposidephosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide;floxuridine; fludarabine phosphate; fluorouracil; fluorocitabine;fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride;hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine;interleukin II (including recombinant interleukin II, or rIL2),interferon alfa-2a; interferon alfa-2b; interferon alfa-n1; interferonalfa-n3; interferon beta-I a; interferon gamma-I b; iproplatin;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ormaplatin; oxisuran; pegaspargase;peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman;piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimersodium; porfiromycin; prednimustine; procarbazine hydrochloride;puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide;safingol; safingol hydrochloride; semustine; simtrazene; sparfosatesodium; sparsomycin; spirogermanium hydrochloride; spiromustine;spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin;tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin;teniposide; teroxirone; testolactone; thiamiprine; thioguanine;thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestoloneacetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate;triptorelin; tubulozole hydrochloride; uracil mustard; uredepa;vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate;vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride.

Other drugs that can be used in combination with the compounds of theinvention to treat a patient with a proliferative disorder such ascancer, or to prevent the reoccurrence of a proliferative disorder suchas cancer, include, but are not limited to: 20-epi-1,25 dihydroxyvitaminD3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron;doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen;ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur;epirubicin; epristeride; estramustine analogue; estrogen agonists;estrogen antagonists; etanidazole; etoposide phosphate; exemestane;fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone BI; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofuran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer. Preferred additional anti-cancer drugs are 5-fluorouraciland leucovorin.

Examples of therapeutic antibodies that can be used in combination withthe compounds of the invention to treat a proliferative disorder such ascancer, or to prevent the reoccurrence of a proliferative disorder suchas cancer, include but are not limited to HERCEPTIN® (Trastuzumab)(Genentech, California) which is a humanized anti-HER2 monoclonalantibody for the treatment of patients with metastatic breast cancer;REOPRO® (abciximab) (Centocor) which is an anti-glycoprotein IIb/IIIareceptor on the platelets for the prevention of clot formation; ZENAPAX®(daclizumab) (Roche Pharmaceuticals, Switzerland) which is animmunosuppressive, humanized anti-CD25 monoclonal antibody for theprevention of acute renal allograft rejection; PANOREX™ which is amurine anti-17-IA cell surface antigen IgG2a antibody (GlaxoWellcome/Centocor); BEC2 which is a murine anti-idiotype (GD3 epitope)IgG antibody (ImClone Systems); IMC-C225 which is a chimeric anti-EGFRIgG antibody (ImClone Systems); VITAXINT™ which is a humanized anti-αVβ3integrin antibody (Applied Molecular Evolution/MedImmune); Campath1H/LDP-03 which is a humanized anti CD52 IgG1 antibody (Leukosite);Smart M195 which is a humanized anti-CD33 IgG antibody (Protein DesignLab/Kanebo); RITUXAN™ which is a chimeric anti-CD20 IgG1 antibody (IDECPharm/Genentech, Roche/Zettyaku); LYMPHOCIDE™ which is a humanizedanti-CD22 IgG antibody (Immunomedics); LYMPHOCIDE™ Y-90 (Immunomedics);Lymphoscan (Tc-99m-labeled; radioimaging; Immunomedics); Nuvion (againstCD3; Protein Design Labs); CM3 is a humanized anti-ICAM3 antibody (ICOSPharm); IDEC-114 is a primatied anti-CD80 antibody (IDECPharm/Mitsubishi); ZEVALIN™ is a radiolabelled murine anti-CD20 antibody(IDEC/Schering AG); IDEC-131 is a humanized anti-CD40L antibody(IDEC/Eisai); IDEC-151 is a primatized anti-CD4 antibody (IDEC);IDEC-152 is a primatized anti-CD23 antibody (IDEC/Seikagaku); SMARTanti-CD3 is a humanized anti-CD3 IgG (Protein Design Lab); 5G1.1 is ahumanized anti-complement factor 5 (C5) antibody (Alexion Pharm); D2E7is a humanized anti-TNF-α antibody (CAT/BASF); CDP870 is a humanizedanti-TNF-α Fab fragment (Celltech); IDEC-151 is a primatized anti-CD4IgG1 antibody (IDEC Pharm/SmithKline Beecham); MDX-CD4 is a humananti-CD4 IgG antibody (Medarex/Eisai/Genmab); CD20-sreptdavidin(+biotin-yttrium 90; NeoRx); CDP571 is a humanized anti-TNF-α IgG4antibody (Celltech); LDP-02 is a humanized anti-α4β7 antibody(LeukoSite/Genentech); OrthoClone OKT4A is a humanized anti-CD4 IgGantibody (Ortho Biotech); ANTOVA™ is a humanized anti-CD40L IgG antibody(Biogen); ANTEGREN™ is a humanized anti-VLA-4 IgG antibody (Elan); andCAT-152 is a human anti-TGF-β2 antibody (Cambridge Ab Tech).

Chemotherapeutic agents that can be used in combination with thecompounds of the invention to treat a patient with a proliferativedisorder such as cancer, or to prevent the reoccurrence of aproliferative disorder such as cancer, include but are not limited toalkylating agents, antimetabolites, natural products, or hormones.Examples of alkylating agents useful for the treatment or prevention ofT-cell malignancies in the methods and compositions of the inventioninclude but are not limited to, nitrogen mustards (e.g.,mechloroethamine, cyclophosphamide, chlorambucil, etc.), alkylsulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne,etc.), or triazenes (decarbazine, etc.). Examples of antimetabolitesuseful for the treatment or prevention of T-cell malignancies in themethods and compositions of the invention include but are not limited tofolic acid analog (e.g., methotrexate), or pyrimidine analogs (e.g.,Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine,pentostatin). Examples of natural products useful for the treatment orprevention of T-cell malignancies in the methods and compositions of theinvention include but are not limited to vinca alkaloids (e.g.,vinblastin, vincristine), epipodophyllotoxins (e.g., etoposide),antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g.,L-asparaginase), or biological response modifiers (e.g., interferonalpha).

Examples of alkylating agents useful for the treatment or prevention ofa proliferative disorder such as cancer in the methods and compositionsof the invention include but are not limited to, nitrogen mustards(e.g., mechloroethamine, cyclophosphamide, chlorambucil, melphalan,etc.), ethylenimine and methylmelamines (e.g., hexamethlymelamine,thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine, lomusitne, semustine, streptozocin, etc.), or triazenes(decarbazine, etc.). Examples of antimetabolites useful for thetreatment or prevention of cancer in the methods and compositions of theinvention include but are not limited to folic acid analog (e.g.,methotrexate), or pyrimidine analogs (e.g., fluorouracil, floxouridine,Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine,pentostatin). Examples of natural products useful for the treatment orprevention of cancer in the methods and compositions of the inventioninclude but are not limited to vinca alkaloids (e.g., vinblastin,vincristine), epipodophyllotoxins (e.g., etoposide, teniposide),antibiotics (e.g., actinomycin D, daunorubicin, doxorubicin, bleomycin,plicamycin, mitomycin), enzymes (e.g., L-asparaginase), or biologicalresponse modifiers (e.g., interferon alpha). Examples of hormones andantagonists useful for the treatment or prevention of cancer in themethods and compositions of the invention include but are not limited toadrenocorticosteroids (e.g., prednisone), progestins (e.g.,hydroxyprogesterone caproate, megestrol acetate, medroxyprogesteroneacetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol),antiestrogen (e.g., tamoxifen), androgens (e.g., testosteronepropionate, fluoxymesterone), antiandrogen (e.g., flutamide),gonadotropin releasing hormone analog (e.g., leuprolide). Other agentsthat can be used in the methods and with the compositions of theinvention for the treatment or prevention of cancer include platinumcoordination complexes (e.g., cisplatin, carboblatin), anthracenedione(e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methylhydrazine derivative (e.g., procarbazine), adrenocortical suppressant(e.g., mitotane, aminoglutethimide).

In one embodiment, the compounds of the invention can be used incombination with an immunotherapeutic agent for the treatment of aproliferative disorder such as cancer, or to prevent the reoccurrence ofa proliferative disorder such as cancer. Immunotherapy (also calledbiological response modifier therapy, biologic therapy, biotherapy,immune therapy, or biological therapy) is treatment that uses parts ofthe immune system to fight disease. Immunotherapy can help the immunesystem recognize cancer cells, or enhance a response against cancercells. Immunotherapies include active and passive immunotherapies.Active immunotherapies stimulate the body's own immune system whilepassive immunotherapies generally use immune system components createdoutside of the body.

Examples of active immunotherapies include: cancer vaccines, tumor cellvaccines (autologous or allogeneic), viral vaccines, dendritic cellvaccines, antigen vaccines, anti-idiotype vaccines, DNA vaccines,Lymphokine-Activated Killer (LAK) Cell Therapy, or Tumor-InfiltratingLymphocyte (TIL) Vaccine with Interleukin-2 (IL-2). Activeimmunotherapies are currently being used to treat or being tested totreat various types of cancers, including melanoma, kidney (renal)cancer, bladder cancer, prostate cancer, ovarian cancer, breast cancer,colorectal cancer, lung cancer, leukemia, prostate cancer, non-Hodgkin'slymphoma, pancreatic cancer, lymphoma, multiple myeloma, head and neckcancer, liver cancer, malignant brain tumors, and advanced melanoma.

Examples of passive immunotherapies include: monoclonal antibodies andtargeted therapies containing toxins. Monoclonal antibodies includenaked antibodies and conjugated antibodies (also called tagged, labeled,or loaded antibodies). Naked monoclonal antibodies do not have a drug orradioactive material attached whereas conjugated monoclonal antibodiesare joined to a chemotherapy drug (chemolabeled), a radioactive particle(radiolabeled), or a toxin (immunotoxin). A number of naked monoclonalantibody drugs have been approved for treating cancer, including:

Rituximab (Rituxan), an antibody against the CD20 antigen used to treatB cell non-Hodgkin lymphoma; Trastuzumab (Herceptin), an antibodyagainst the HER2 protein used to treat advanced breast cancer;Alemtuzumab (Campath), an antibody against the CD52 antigen used totreat B cell chronic lymphocytic leukemia (B-CLL); Cetuximab (Erbitux),an antibody against the EGFR protein used in combination with irinotecanto treat advanced colorectal cancer and to treat head and neck cancers;and Bevacizumab (Avastin) which is an antiangiogenesis therapy thatworks against the VEGF protein and is used in combination withchemotherapy to treat metastatic colorectal cancer. A number ofconjugated monoclonal antibodies have been approved for treating cancer,including: Radiolabeled antibody Ibritumomab tiuxetan (Zevalin) whichdelivers radioactivity directly to cancerous B lymphocytes and is usedto treat B cell non-Hodgkin lymphoma; radiolabeled antibody Tositumomab(Bexxar) which is used to treat certain types of non-Hodgkin lymphoma;and immunotoxin Gemtuzumab ozogamicin (Mylotarg) which containscalicheamicin and is used to treat acute myelogenous leukemia (AML).BL22 is a conjugated monoclonal antibody currently in testing fortreating hairy cell leukemia and there are several immunotoxin clinicaltrials in progress for treating leukemias, lymphomas, and brain tumors.There are also approved radiolabeled antibodies used to detect cancer,including OncoScint for detecting colorectal and ovarian cancers andProstaScint for detecting prostate cancers. Targeted therapiescontaining toxins are toxins linked to growth factors and do not containantibodies. An example of an approved targeted therapy containing toxinsis denileukin diftitox (Ontak) which is used to treat a type of skinlymphoma (cutaneous T cell lymphoma).

Examples of adjuvant immunotherapies include: cytokines, such asgranulocyte-macrophage colony-stimulating factor (GM-CSF),granulocyte-colony stimulating factor (G-CSF), macrophage inflammatoryprotein (MIP)-1-alpha, interleukins (including IL-1, IL-2, IL-4, IL-6,IL-7, IL-12, IL-15, IL-18, IL-21, and IL-27), tumor necrosis factors(including TNF-alpha), and interferons (including IFN-alpha, IFN-beta,and IFN-gamma); aluminum hydroxide (alum); Bacille Calmette-Guérin(BCG); Keyhole limpet hemocyanin (KLH); Incomplete Freund's adjuvant(IFA); QS-21; DETOX; Levamisole; and Dinitrophenyl (DNP). Clinicalstudies have shown that combining IL-2 with other cytokines, such asIFN-alpha, can lead to a synergistic response.

Several types of immunotherapies are being used to treat melanomapatients. IFN-alpha and IL-2 are approved for treatment of people withmetastatic melanoma. BCG is being tested in combination with melanomavaccines and other immunotherapies. Tumor-infiltrating lymphocytes havebeen shown to shrink melanoma tumors in a phase 1 clinical trial. Humanmonoclonal antibodies to ganglioside antigens have been shown to regresscutaneous recurrent melanoma tumors. Some autologous and allogeneictumor cell vaccines, antigen vaccines (including polyvalent antigenvaccines), and dendritic cell vaccines have also been shown to shrinktumors. Clinical trials continue for these and other melanomaimmunotherapies. Melanoma patients with a high IgM response oftensurvive better than those who elicit no or low IgM antibodies (Morton etal., 1992). Combined IL-12/TNF-alpha immunotherapy has been shown tosignificantly retard tumor growth in three tumor models in mice (B16F10melanoma, Lewis lung (LL/2) carcinoma and LI sarcoma) as compared withcontrols and mice treated with either cytokine alone. IFN-alpha isapproved for the treatment of malignant melanoma, chronic myelogenousleukemia (CML), hairy cell leukemia, and Kaposi's sarcoma.

Several types of immunotherapies are being used to treat patients thathave renal cancer. IFN-alpha and IL-2 are approved for treatment ofpeople with metastatic renal (kidney) cancer. A combination therapyusing IL-2, interferon, and chemotherapy is being tested for treatmentof renal cancer. Treatment with a tumor cell vaccine plus the adjuvantBCG has been shown to shrink tumors in some advanced renal cancerpatients. DNA vaccines and tumor-infiltrating lymphocytes are also beingtested as treatments for renal cancer. Chimeric bispecific G250/anti-CD3monoclonal antibodies have been shown to mediate cell lysis of renalcell carcinoma cell lines by cloned human CD8+ T cells or by IL-2stimulated peripheral blood lymphocytes.

As used herein, a “microtubulin stabilizer” means an anti-cancer agentwhich acts by arresting cells in the G2-M phases due to stabilization ofmicrotubules. Agents which are microtubulin stabilizers can be used incombination with the compounds of the invention to treat patients havinga proliferative disorder such as cancer, or to prevent the reoccurrenceof a proliferative disorder such as cancer. Examples of microtubulinstabilizers include taxol and taxol analogues. Additional examples ofmicrotubulin stabilizers included without limitation the followingmarketed drugs and drugs in development: Discodermolide (also known asNVP-XX-A-296); Epothilones (such as Epothilone A, Epothilone B,Epothilone C (also known as desoxyepothilone A or dEpoA); Epothilone D(also referred to as KOS-862, dEpoB, and desoxyepothilone B); EpothiloneE; Epothilone F; Epothilone B N-oxide; Epothilone A N-oxide;16-aza-epothilone B; 21-aminoepothilone B (also known as BMS-310705);21-hydroxyepothilone D (also known as Desoxyepothilone F and dEpoF),26-fluoroepothilone); FR-182877 (Fujisawa, also known as WS-9885B),BSF-223651 (BASF, also known as ILX-651 and LU-223651); AC-7739(Ajinomoto, also known as AVE-8063A and CS-39.HCl); AC-7700 (Ajinomoto,also known as AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A);Fijianolide B; Laulimalide; Caribaeoside; Caribaeolin; Taccalonolide;Eleutherobin; Sarcodictyin; Laulimalide; Dictyostatin-1; Jatrophaneesters; and analogs and derivatives thereof.

As used herein, a “microtubulin inhibitor” means an anti-cancer agentwhich acts by inhibiting tubulin polymerization or microtubule assembly.Agents which are microtubulin inhibitors can be used in combination withthe compounds of the invention to treat patients having a proliferativedisorder such as cancer, or to prevent the reoccurrence of aproliferative disorder such as cancer. Examples of microtubulininhibitors include without limitation the following marketed drugs anddrugs in development: Erbulozole (also known as R-55104); Dolastatin 10(also known as DLS-10 and NSC-376128); Mivobulin isethionate (also knownas CI-980); Vincristine; NSC-639829; ABT-751 (Abbot, also known asE-7010); Altorhyrtins (such as Altorhyrtin A and Altorhyrtin C);Spongistatins (such as Spongistatin 1, Spongistatin 2, Spongistatin 3,Spongistatin 4, Spongistatin 5, Spongistatin 6, Spongistatin 7,Spongistatin 8, and Spongistatin 9); Cemadotin hydrochloride (also knownas LU-103793 and NSC-D-669356); Auristatin PE (also known asNSC-654663); Soblidotin (also known as TZT-1027), LS-4559-P (Pharmacia,also known as LS-4577); LS-4578 (Pharmacia, also known as LS-477-P);LS-4477 (Pharmacia), LS-4559 (Pharmacia); RPR-112378 (Aventis);Vincristine sulfate; DZ-3358 (Daiichi); GS-164 (Takeda); GS-198(Takeda); KAR-2 (Hungarian Academy of Sciences); SAH-49960(Lilly/Novartis); SDZ-268970 (Lilly/Novartis); AM-97 (Armad/KyowaHakko); AM-132 (Armad); AM-138 (Armad/Kyowa Hakko); IDN-5005 (Indena);Cryptophycin 52 (also known as LY-355703); Vitilevuamide; Tubulysin A;Canadensol; Centaureidin (also known as NSC-106969); T-138067 (Tularik,also known as T-67, TL-138067 and TI-138067); COBRA-1 (Parker HughesInstitute, also known as DDE-261 and WHI-261); H10 (Kansas StateUniversity); H16 (Kansas State University); Oncocidin A 1 (also known asBTO-956 and DIME); DDE-313 (Parker Hughes Institute); SPA-2 (ParkerHughes Institute); SPA-1 (Parker Hughes Institute, also known asSPIKET-P); 3-IAABU (Cytoskeleton/Mt. Sinai School of Medicine, alsoknown as MF-569); Narcosine (also known as NSC-5366); Nascapine, D-24851(Asta Medica), A-105972 (Abbott); Hemiasterlin; 3-BAABU(Cytoskeleton/Mt. Sinai School of Medicine, also known as MF-191); TMPN(Arizona State University); Vanadocene acetylacetonate; T-138026(Tularik); Monsatrol; Inanocine (also known as NSC-698666); 3-IAABE(Cytoskeleton/Mt. Sinai School of Medicine); A-204197 (Abbott); T-607(Tularik, also known as T-900607); RPR-115781 (Aventis); Eleutherobins(such as Desmethyleleutherobin, Desaetyleleutherobin, Isoeleutherobin A,and Z-Eleutherobin); Halichondrin B; D-64131 (Asta Medica); D-68144(Asta Medica); Diazonamide A; A-293620 (Abbott); NPI-2350 (Nereus);TUB-245 (Aventis); A-259754 (Abbott); Diozostatin; (−)-Phenylahistin(also known as NSCL-96F037); D-68838 (Asta Medica); D-68836 (AstaMedica); Myoseverin B; D-43411 (Zentaris, also known as D-81862);A-289099 (Abbott); A-318315 (Abbott); HTI-286 (also known as SPA-110,trifluoroacetate salt) (Wyeth); D-82317 (Zentaris); D-82318 (Zentaris);SC-12983 (NCl); Resverastatin phosphate sodium; BPR-0Y-007 (NationalHealth Research Institutes); SSR-250411 (Sanofi); Combretastatin A4; andanalogs and derivatives thereof.

Taxol®, also referred to as “paclitaxel”, is a well-known anti-cancerdrug which acts by enhancing and stabilizing microtubule formation. Thestructure of Taxol® is shown in FIG. 1. Many analogs of Taxol® areknown, including Taxotere®, the structure of which is shown in FIG. 2.Taxotere® is also referred to as “docetaxol”. The structures of otherTaxol® analogs are shown in FIGS. 3-23. These compounds have the basictaxane skeleton as a common structure feature and have also been shownto have the ability to arrest cells in the G2-M phases due tostabilization of microtubules. Thus, it is apparent from FIGS. 3-23 thata wide variety of substituents can decorate the taxane skeleton withoutadversely affecting biological activity. It is also apparent that zero,one or both of the cyclohexane rings of a taxol analog can have a doublebond at the indicated positions. For clarity purposes, the basic taxaneskeleton is shown below in Structural Formula (XXVII):

Double bonds have been omitted from the cyclohexane rings in the taxaneskeleton represented by Structural Formula (XXVII). The basic taxaneskeleton can include zero or one double bond in one or both cyclohexanerings, as indicated in FIGS. 3-23 and Structural Formulas (XXVIII) and(XXIX) below. A number of atoms have also been omitted from StructuralFormula (XXVII) to indicate sites in which structural variation commonlyoccurs among taxol analogs. For example, substitution on the taxaneskeleton with simply an oxygen atom indicates that hydroxyl, acyl,alkoxy or another oxygen-bearing substituent is commonly found at thesite. These and other substitutions on the taxane skeleton can be madewithout losing the ability to enhance and stabilize microtubuleformation. Thus, the term “Taxol® analog” is defined herein to mean acompound which has the basic taxane skeleton and which promotesmicrotubule formation. Taxol® analogs may be formulated as ananoparticle colloidal composition to improve the infusion time and toeliminate the need to deliver the drug with Cremophor which causeshypersensitivity reactions in some patients. An example of a Taxol®analog formulated as a nanoparticle colloidal composition is Abraxanewhich is a nanoparticle colloidal composition of protein-stabilizedpaclitaxel that is reconstituted in saline.

Typically, the Taxol® analogs used herein are represented by StructuralFormula (XXVIII) or (XXIX):

R₁₀ is a lower alkyl group, a substituted lower alkyl group, a phenylgroup, a substituted phenyl group, —SR₁₉, —NHR₁₉ or —OR₁₉.

R₁₁ is a lower alkyl group, a substituted lower alkyl group, an arylgroup or a substituted aryl group.

R₁₂ is —H, —OH, lower alkyl, substituted lower alkyl, lower alkoxy,substituted lower alkoxy, —O—C(O)-(lower alkyl), —O—C(O)-(substitutedlower alkyl), —O—CH₂—O-(lower alkyl)-S—CH₂—O-(lower alkyl).

R₁₃ is —H, —CH₃, or, taken together with R₁₄, —CH₂—.

R₁₄ is —H, —OH, lower alkoxy, —O—C(O)-(lower alkyl), substituted loweralkoxy, —O—C(O)-(substituted lower alkyl), —O—CH₂—O—P(O)(OH)₂,—O—CH₂—O-(lower alkyl), —O—CH₂—S-(lower alkyl) or, taken together withR₂₀, a double bond.

R₁₅—H, lower acyl, lower alkyl, substituted lower alkyl, alkoxymethyl,alkthiomethyl, —OC(O)—O(lower alkyl), —OC(O)—O(substituted lower alkyl),—OC(O)—NH(lower alkyl) or —OC(O)—NH(substituted lower alkyl).

R₁₆ is phenyl or substituted phenyl.

R₁₇ is —H, lower acyl, substituted lower acyl, lower alkyl, substituted,lower alkyl, (lower alkoxy)methyl or (lower alkyl)thiomethyl.

R₁₈—H, —CH₃ or, taken together with R₁₇ and the carbon atoms to whichR₁₇ and R₁₈ are bonded, a five or six membered a non-aromaticheterocyclic ring.

R₁₉ is a lower alkyl group, a substituted lower alkyl group, a phenylgroup, a substituted phenyl group.

R₂₀ is —H or a halogen.

R₂₁ is —H, lower alkyl, substituted lower alkyl, lower acyl orsubstituted lower acyl.

Preferably, the variables in Structural Formulas (XXVIII) and (XXIX) aredefined as follows: R₁₀ is phenyl, tert-butoxy, —S—CH₂—CH—(CH₃)₂,—S—CH(CH₃)₃, —S—(CH₂)₃CH₃, —O—CH(CH₃)₃, —NH—CH(CH₃)₃, —CH═C(CH₃)₂ orpara-chlorophenyl; R₁₁ is phenyl, (CH₃)₂CHCH₂—, -2-furanyl, cyclopropylor para-toluoyl; R₁₂ is —H, —OH, CH₃CO— or —(CH₂)₂—N-morpholino; R₁₃ ismethyl, or, R₁₃ and R₁₄, taken together, are —CH₂—;

R₁₄ is —H, —CH₂SCH₃ or —CH₂—O—P(O)(OH)₂; R₁₅ is CH₃CO—;

R₁₆ is phenyl; R₁₇—H, or, R₁₇ and R₁₈, taken together, are —O—CO—O—;

R₁₈ is —H; R₂₀ is —H or —F; and R₂₁ is —H, —C(O)—CHBr—(CH₂)₁₃—CH₃ or—C(O)—(CH₂)₁₄—CH₃; —C(O)—CH₂—CH(OH)—COOH,—C(O)—CH₂—O—C(O)—CH₂CH(NH₂)—CONH₂, —C(O)—CH₂—O—CH₂CH₂OCH₃ or—C(O)—O—C(O)—CH₂CH₃.

A Taxol® analog can also be bonded to or be pendent from apharmaceutically acceptable polymer, such as a polyacrylamide. Oneexample of a polymer of this type is shown in FIG. 24. The term “Taxol®analog”, as it is used herein, includes such polymers.

In some embodiments, Taxol® analogs have a taxane skeleton representedby Structural Formula XXX, wherein Z is O, S, or NR. Taxol® analogs thathave the taxane skeleton shown in Structural Formula XXX can havevarious substituents attached to the taxane skeleton and can have adouble bond in zero, one or both of the cyclohexane rings as shown, forexample in FIGS. 3-23.

Various Taxol® analogs and Taxol® formulations are described inHennenfent et al. (2006) Annals of Oncology 17:735-749; Gradishar (2006)Expert Opin. Pharmacother. 7(8): 1041-53; Attard et al. (2006) PatholBiol 54(2):72-84; Straubinger et al. (2005) Methods Enzymol. 391:97-117;Ten Tije et al. (2003) Clin Pharmacokinet. 42(7):665-85; and Nuijen etal. (2001) Invest New Drugs. 19(2):143-53, the entire teachings of whichare incorporated herein by reference.

In some embodiments, the invention provides a method for treating orinhibiting angiogenesis in a subject in need thereof, comprisingadministering to the subject an effective amount of a compoundrepresented by formula (I) through (XXVI) and Table 1. As used herein,the term “angiogenesis” refers to a fundamental process of generatingnew blood vessels in tissues or organs. Angiogenesis is involved with orassociated with many diseases or conditions, including, but not limitedto: cancer; ocular neovascular disease; age-related maculardegeneration; diabetic retinopathy, retinopathy of prematurity; cornealgraft rejection; neovascular glaucoma; retrolental fibroplasias;epidemic keratoconjunctivitis; Vitamin A deficiency; contact lensoverwear; atopic keratitis; superior limbic keratitis; pterygiumkeratitis sicca; sjogrens; acne rosacea; warts; eczema; phylectenulosis;syphilis; Mycobacteria infections; lipid degeneration; chemical burns;bacterial ulcers; fungal ulcers; Herpes simplex infections; Herpeszoster infections; protozoan infections; Kaposi's sarcoma; Mooren'sulcer; Terrien's marginal degeneration; marginal keratolysis; rheumatoidarthritis; systemic lupus; polyarteritis; trauma; Wegener's sarcoidosis;scleritis; Stevens-Johnson disease; pemphigoid; radial keratotomy;corneal graph rejection; diabetic retinopathy; macular degeneration;sickle cell anemia; sarcoid; syphilis; pseudoxanthoma elasticum; Paget'sdisease; vein occlusion; artery occlusion; carotid obstructive disease;chronic uveitis/vitritis; mycobacterial infections; Lyme's disease;systemic lupus erythematosis; retinopathy of prematurity; Eales'disease; Behcet's disease; infections causing a retinitis orchoroiditis; presumed ocular histoplasmosis; Best's disease; myopia;optic pits; Stargardt's disease; pars planitis; chronic retinaldetachment; hyperviscosity syndromes; toxoplasmosis; trauma andpost-laser complications; diseases associated with rubeosis(neovasculariation of the angle); diseases caused by the abnormalproliferation of fibrovascular or fibrous tissue including all forms ofproliferative vitreoretinopathy; rheumatoid arthritis; osteoarthritis;ulcerative colitis; Crohn's disease; Bartonellosis; atherosclerosis;Osler-Weber-Rendu disease; hereditary hemorrhagic telangiectasia;pulmonary hemangiomatosis; pre-eclampsia; endometriosis; fibrosis of theliver and of the kidney; developmental abnormalities (organogenesis);skin disclolorations (e.g., hemangioma, nevus flammeus, or nevussimplex); wound healing; hypertrophic scars, i.e., keloids; woundgranulation; vascular adhesions; cat scratch disease (Rochele ninaliaquintosa); ulcers (Helicobacter pylori); keratoconjunctivitis;gingivitis; periodontal disease; epulis; hepatitis; tonsillitis;obesity; rhinitis; laryngitis; tracheitis; bronchitis; bronchiolitis;pneumonia; interstitial pulmonary fibrosis; pulmonary edema;neurodermitis; thyroiditis; thyroid enlargement; endometriosis;glomerulonephritis; gastritis; inflammatory bone and cartilagedestruction; thromboembolic disease; and Buerger's disease.Anti-angiogenesis can be demonstrated by any method known to thoseskilled in the art, such as the method described herein in Examples 10and 11.

Anti-angiogenesis agents that can be co-administered with the compoundsof the invention include Dalteparin, Suramin, ABT-510, Combretastatin A4Phosphate, Lenalidomide, LY317615 (Enzastaurin), Soy Isoflavone(Genistein; Soy Protein Isolate), Thalidomide, AMG-706, Anti-VEGFAntibody (Bevacizumab; Avastin™), AZD2171, Bay 43-9006 (Sorafenibtosylate), PI-88, PTK787/ZK 222584 (Vatalanib), SUI 1248 (Sunitinibmalate), VEGF-Trap, XL184, ZD6474, ATN-161, EMD 121974 (Cilenigtide),Celecoxib, Angiostatin, Endostatin, Regranex, Apligraf, Paclitaxel,tetracyclines, clarithromycin, lasix, captopril, aspirin, Vitamin D3analogs, retinoids, Imiquomod, Interferon alfa2a, Minocycline, copperpeptide containing dressings, Lucentis™, ATG002, Pegaptanib Sodium,Tryptophanyl-tRNA synthetase, squalamine lactate, anecortave acetate,AdPEDF, AG-013958, JSM6427, TG100801, Veglin, ascorbic acid ethers (andtheir analogs), and Pamidronate.

The compounds of the invention can be prepared by the schemes below orby any method known to those in the art.

Compounds of the invention can also be prepared according to U.S. Pat.No. 6,800,660, U.S. Pat. No. 6,762,204, U.S. Pat. No. 6,825,235, orPatent Appln. No. PCT/US2007/018378, which are incorporated by referenceherein in their entirety.

The entire teachings of each of the references cited in the aboveschemes are incorporated herein by reference.

The present invention is illustrated by the following examples, whichare not intended to be limiting in any way.

EXEMPLIFICATION Example 1 Compound 1:2-((dimethylamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

To a solution of B (1.0 mmol) in THF (5 ml), was addedN,N-dimethylformamide dimethyl acetal (2.0 mmol). The reaction mixturewas stirred at RT for 2 hr. Completion was judged by TLC. The reactionmixture was diluted with water (2 ml) and extracted with CH₂Cl₂ (2×10ml) and the organic phase washed with water and brine, dried (Na₂SO₄),filtered, evaporated and purified by flash chromatography on a column ofsilica gel (2-5% MeOH in CH₂Cl₂) and to give Compound 1 (365 mg) in 80%yield.

ESMS clcd for C₂₂H₂₅N₅O₂S₂: 455.14. Found: 456.2 (M+H)⁺.

Compound B can be prepared according to U.S. Pat. No. 6,800,660 and U.S.Pat. No. 6,825,235, which are incorporated by reference herein in theirentirety.

To a solution of compound Compound 1 (1.0 mmol) in dioxane (10 ml), wasadded the amine (2.5 mmol). The reaction mixture was stirred at RT for3-5 hr. Completion was judged by TLC. The reaction mixture was dilutedwith CH₂Cl₂ (20 ml) and the organic phase washed with water and brine,dried (Na₂SO₄), filtered, evaporated. The residue was purified by thecolumn chromatography (silica gel 2-5% MeOH in CH₂Cl₂) to give productA.

Compound 4:2-((2-hydroxyethylamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₂H₂₅N₅O₃S₂: 471.14. Found: 472.2 (M+H)⁺.

Compound 5:2-((2-methoxyethylamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₃H₂₇N₅O₃S₂: 485.16. Found: 486.1 (M+H)⁺.

Compound 6:2-((methoxyamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₁H₂₃N₅O₃S₂: 457.12. Found: 458.2 (M+H)⁺.

Compound 7:2-((2,2-dimethylhydrazinyl)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₂H₂₆N₆O₂S₂: 470.16. Found: 471.2 (M+H)⁺.

Compound 8:2-(aminomethylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₀H₂₁N₅O₂S₂: 427.11. Found: 428.2 (M+H)⁺.

Compound 9:N′1,N′3-dimethyl-2-((methylamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₁H₂₃N₅O₂S₂: 441.13. Found: 442.2 (M+H)⁺.

Compound 10:2-((cyclopropylamino)methylene)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₃H₂₅N₅O₂S₂: 467.14. Found: 468.2 (M+H)⁺.

Compound 11:N′1,N′3-dimethyl-2-((morpholinoamino)methylene)-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₄H₂₈N₆O₃S₂: 512.17. Found: 513.2 (M+H)⁺.

To a solution of B (400 mg, 1.0 mmol) in THF (5 ml) at 0° C. was addedtert-butyl nitrite (105 mg, 1.1 mmol) and hydrogen chloride 2.0 Msolution in diethyl ether (0.7 ml, 1.4 mmol). The reaction mixture wasstirred at 0° C. for 30 min. and then the solution was allowed to reachRT. Completion was judged by TLC. The reaction mixture was diluted withwater (2 ml) and extracted with CH₂Cl₂ (2×10 ml) and the organic phasewashed with water and brine, dried (Na₂SO₄), filtered, evaporated andpurified by flash chromatography on a column of silica gel (1:1Hexane:EtOAc) and to give Compound 2 (325 mg) in 75% yield.

To a solution of Compound 2 (220 mg, 0.5 mmol) in CH₂Cl₂ (10 ml) wascarefully added solution CH₂N₂ in diethyl ether at RT, (TLC control).When the reaction is finish, the solvent was removed under reducedpressure and the product was purified by flash chromatography on acolumn of silica gel (1:1 Hexane:EtOAc) and to give Compound 3 (175 mg)in 79% yield.

Compound 2:2-(hydroxyimino)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₁₉H₁₉N₅O₃S₂: 429.09. Found: 430.1 (M+H)⁺.

Compound 3:2-(methoxyimino)-N′1,N′3-dimethyl-N′1,N′3-di(phenylcarbonothioyl)malonohydrazide

ESMS clcd for C₂₀H₂₁N₅O₃S₂: 443.11. Found: 444.2 (M+H)⁺.

Example 2 Compounds of the Invention Enhance the Anti-Cancer Activity ofAnti-Cancer Agents In Vivo

A. General Procedure of In Vivo Anti-Tumor Study

The in vivo anti-cancer enhancing effect of novel compounds are assessedin tumor bearing mice using the tumor growth inhibition assay. Tumorcells are implanted by injection of a tumor cell suspensionsubcutaneously in the flank of a mouse. Treatment of the tumor with acompound of the invention and another anti-cancer agent (e.g.,paclitaxel, which will be used hereinafter by way of example) is begunafter the tumor had been established (volume was about 100 mm³). Theanimals are then started on a multiple injection schedule where thecompound and paclitaxel are given by IV route of administration. Tumorsare measured two times a week. During the course of this assay, animalsare monitored daily for signs of toxicity including body weight loss.

B. Procedure

A supplemented media is prepared from 50% DMEM/Dulbecco Modified EagleMedium (High Glucose), 50% RPMI 1640, 10% FBS/Fetal Bovine Serum(Hybridoma Tested; Sterile Filtered), 1% L-Glutamine, 1%Penicillin-Streptomycin, 1% MEM Sodium Pyruvate and 1% MEM Non-EssentialAmino Acids. FBS is obtained from Sigma Chemical Co. and otheringredients are obtained from Invitrogen Life Technologies, USA). Thesupplemental Media is warmed to 37° C. and 50 mL of media is added to a175 cm² tissue culture flask.

The cells used in the assay are MDA-435 Human Breast Carcinoma from theAmerican Type Culture Collection. 1 vial of MDA-435 cells from theliquid nitrogen frozen cell stock is removed. The frozen vial of cellsis immediately placed into a 37° C. water bath and gently swirled untilthawed. The freeze-vial is wiped with 70% ethanol and cells areimmediately pipetted into the 175 cm² tissue culture flask containingsupplemented media. The cells are incubated overnight and the media isremoved and replaced with fresh supplemented media the next day. Theflask is incubated until flask became about 90% confluent. Thisgenerally takes anywhere from 5-7 days.

The flask is washed with 10 ml of sterile room temperature phosphatebuffered saline (PBS). The cells are trypsinized by adding 5 ml ofwarmed Trypsin-EDTA (Invitrogen) to the flask of cells. The cells arethen incubated for 2-3 minutes at 37° C. until cells begin to detachfrom the surface of the flask. An equal volume of supplemented media (5ml) is added to the flask. All the cells are collected into 50 ml tube,and centrifuged at 1000 RPM for 5 minutes at 20° C. The supernatant isaspirated and the cell pellet is resuspended in 10 ml of supplementedmedia and the cells are counted. 1-3 million cells/flask are seeded into5-7 tissue culture flasks (175 cm²). Each flask should contain 50 ml ofsupplemented media. The flasks are incubated until about 90% confluent.The passaging of the cells is repeated until enough cells have beengrown for tumor implantation.

The above procedure for trypsinizing and centrifuging the cells arefollowed. The supernatant is aspirated and the cell pellet isresuspended in 10 ml of sterile PBS and the cells are counted. The cellsare centrifuged and then resuspended with appropriate volume of sterilePBS for injection of correct number of cells needed for tumorimplantation. In the case of MDA-435, 100 million cells are suspendedwith 2.0 ml of sterile PBS to a final concentration of 50 millioncells/ml in order to inject 5 million cells in 0.1 ml/mouse.

Mice (CD-1 nu/nu) are obtained from Charles River Laboratories:nomenclature: Crl:CD-1-nuBR, Age: 6-8 weeks. The mice are allowed toacclimate for 1 week prior to their being used in an experimentalprocedure.

Implantation of the MDA-435 tumor cell suspension is generally into thecorpus adiposum of the female CD-1 nu/nu mouse. This fat body is locatedin the ventral abdominal viscera of the mouse. Tumor cells are implantedsubcutaneously into the fat body located in the right quadrant of theabdomen at the juncture of the os coxae (pelvic bone) and the os femoris(femur). 5 million MDA-435 cells in 0.1 ml of sterile PBS are injectedusing 27 G (½ inch) needle. MDA-435 tumors are typically developed 2-3weeks after implantation.

Compound stock solutions are prepared by dissolving the compound incell-culture-grade DMSO (dimethyl sulfoxide) at the desiredconcentration. This stock solution in DMSO is sonicated in an ultrasonicwater bath until all the powder dissolved.

The Formulation Solvent is prepared as follows: 20% of Cremophor RH40(Polyoxyl 40 Hydrogenated Castor Oil obtained from BASF corp.) in wateris prepared by first heating 100% Cremophor RH40 in a water bath at50-60° C. until it liquefied and became clear. 10 ml of the 100%Cremophor RH40 is aliquotted into a conical centrifuge tube containing40 ml of sterile water (1:5 dilution of Cremophor RH40). The 20%Cremophor RH40 solution is reheated until it became clear again, andmixed by inverting the tube several times. This 20% Cremophor RH40solution is stored at room temperature, and was kept for up to 3 months.

Preparation of Dosing Solution for Compound Administration: the compoundstock solution is diluted 1:10 with 20% Cremophor RH40:1) 2.0 ml of 10mg/ml dosing solution of a compound of the invention is prepared bydiluting 100 mg/ml Compound Stock solution with 1.8 ml of 20% CremophorRH40 water solution; and 2) a dosing solution comprising 2.0 ml of 1mg/ml of Paclitaxel (obtained from Sigma Chemical Co.) and 5 mg/ml ofCompound (I) is obtained by mixing 0.1 ml of a compound of the inventionDMSO stock solution (50 mg/ml) and 0.1 ml of Paclitaxel DMSO stocksolution (10 mg/ml) and diluting with 1.8 ml of 20% Cremophor RH40 watersolution. The final formulation for the dosing solution was 10% DMSO,18% Cremophor RH40 and 72% water.

The Dosing Solution (Dosing Volume: 0.01 ml/gram=10 ml/kg) is injectedintravenously into the mice bearing MDA-435 human breast tumor. Thetable below shows a typical dosing protocol

Group Compounds Dosed 1 Vehicle only 2 Paclitaxel (5 mg/kg) 3 Compoundof the invention (50 mg/kg) 4 Paclitaxel (5 mg/kg) and Compound of theinvention (25 mg/kg) 5 Paclitaxel (5 mg/kg) and Compound of theinvention (50 mg/kg)

Results

The compounds of the invention are expected to significantly enhancedanti-tumor activity of Paclitaxel without increasing toxicity.

Examples 3-7

Heat shock proteins (Hsp) are induced under a variety of stressconditions and bind to other proteins to prevent their denaturation.Hsps can protect the cell from apoptotic death. Agents that induce theproduction of Hsp70 can have protective activity against a wide range ofinsults, and may have particular utility in neurological disorders. Theneuroprotectant activity of Hsp70 inducing compounds of the inventioncan be assessed in a variety of animal neurological disease models.Specifically, animal models of stroke, amyotrophic lateral sclerosis,Huntington's disease, Parkinson's disease, and Alzheimer's disease areappropriate settings for testing efficacy. Some example animal modelsare provided below.

Example 3 Cerebral Ischemia (Stroke)

The benefit of the disclosed treatment with Hsp70 inducing compounds ofthe invention can be assessed in rodent models of stroke. For examplethe stroke model described in Longa, et al. (Longa, E. Z., Weinstein, P.R., Carlson, S., and Cummins, R. (1989) Reversible middle cerebralartery occlusion without craniectomy in rats. Stroke 20:84-91) can beutilized.

Rats are anesthetized with ketamine, and then infarction is induced byextracranial vascular occlusion. A 4-0 nylon intraluminal suture isplaced into the cervical internal carotid artery and is advancedintracranially to block blood flow into the middle cerebral artery.Collateral blood flow is reduced by interrupting all branches of theexternal carotid artery and all extracranial branches of the internalcarotid artery. A compounds of the invention can be dosed just prior toor just after induction of the infarction. The dose may be, for example,10 to 100 mg/kg body weight administered once per week, three times perweek, or daily by any conventional mode of administration, e.g., orallyor intravenously. Neurologic deficit, mortality, gross pathology(infarction size), and histochemical staining can be analyzed to assessefficacy of the compounds. Since this is a very acute model, and deathis often observed by three days after infarction, the modeling mayconsist of only a single administration of drug.

Example 4 Familial Amyotrophic Lateral Sclerosis (ALS)

The efficacy of compounds of the invention in the treatment of ALS canbe modeled using the SOD1 transgenic mouse model (Gurney, M. E., Pu, H.,Chiu, A. Y., Dal Canto, M. C., Polchow, C. Y., Alexander, D. D.,Caliendo, J., Hentati, A., Kwon, Y. W., and Deng, H. X. (1994) Motorneuron degeneration in mice that express a human CuZn superoxidedismutase mutation. Science 264:1772-1775). Mutations of human CuZnsuperoxide dismutase (SOD) are found in patients with familial ALS.Expression of the human SOD gene containing a substitution ofglycine-to-alanine at amino acid 93 leads to motor neuron disease intransgenic mice. As a result of motor neuron loss from the spinal cord,the mice became paralyzed and die by 5 to 6 months of age.

To test the efficacy of the Hsp70 inducing compounds of the invention,transgenic mice having the SOD1 mutation (SOD1^(G93A)) are treated withthe compounds, and the effect on disease is monitored. The symptoms areclinically apparent in these animals at 2.5 to 3 months of age.Compounds can be dosed starting at this time. The dose may be, forexample, 10 to 100 mg/kg body weight administered once per week or threetimes per week by the oral or intravenous route. Endpoints includefunctional impairment of motor function as well as histological changes.The latter endpoints include histopathology of brain and spinal cordassessing degeneration of motor neurons and the appearance ofneurofilament-rich inclusions in spinal motor neurons. If long-termadministration is performed, the impact on mouse survival can beassessed.

Example 5 Huntington's Disease (HD)

A transgenic mouse model of HD exists, allowing the testing of Hsp70inducing compounds of the invention for efficacy in this disease setting(Mangiarini, L., Sathasivam, K., Seller, M., Cozens, B., Harper, A.,Hetherington, C., Lawton, M., Trottier, Y., Lehrach, H., Davies, S. W.,and Bates, G. P. (1996) Exon 1 of the HD gene with an expanded CAGrepeat is sufficient to cause a progressive neurological phenotype intransgenic mice. Cell 87:493-506; Carter, R. J., Lione, L. A., Humby,T., Mangiarini, L., Mahal, A., Bates, G. P., Dunnett, S. B., and Morton,A. J. (1999) Characterization of progressive motor deficits in micetransgenic for the human Huntington's disease mutation. J. Neuroscience19:3248-3257). HD is caused by a CAG/polyglutamine repeat expansion.These transgenic mice (R6/2 transgenics) have the 5′ end of the human HDgene with (CAG)115-(CAG)150 repeat expansions. The mice exhibitprogressive neurological pathologies similar to HD, including abnormaland involuntary movements, tremors, and epileptic seizures.

These transgenic mice show overt behavioral changes at approximately 8weeks of age. As early as 5 to 6 weeks of age, they display more subtledeficiencies in motor skills. Hsp70 inducing compounds of the inventioncan be administered by intravenous or oral administration at doses of10-100 mg per kg of body weight starting at various times (for example,at 5 to 6 weeks of age). Compounds can be given on multiple differentdosing schedules (e.g., once per week versus three times per week).Performance on one or more rodent motor tests such as swimming tank,beam walking, rotarod apparatus, and footprint test (see Carter, et al.,1999) can be performed to assess the activity of the compounds inpreventing loss of neurological function in HD mice.

Example 6 Parkinson's Disease (PD)

There are two widely employed models of PD in which disease is inducedby chemical treatment. These are the 6-OHDA (Zigmond, M. J. andStricker, E. M. (1984) Parkinson's disease: studies with an animalmodel. Life Sci. 35:5-18; Sauer, H. and Oertel, W. H. (1994) Progressivedegeneration of nigrostriatal dopamine neurons following intrastriatalterminal lesions with 6-hydroxydopamine: a combined retrograde tracingand immunocytochemical study in the rat. Neuroscience 59:401-415) andthe MPTP (Langston, J. W., Formo, L. S., Rebert, C. S., and Irwin, I.(1984) Selective nigral toxicity after systemic administration of1-methyl-4-phenyl-1,2,5,6-tetrahydropyrine (MPTP) in the squirrelmonkey. Brain Res. 292:390-4) models. An example of a test of Hsp70inducing compounds of the invention using the 6-OHDA is described.

Young adult male rats are injected with Fluoro-Gold (FG) by stereotacticinjection into the striatum in the brain in order to facilitatevisualization of the neurons in the substantia nigra, the site of PD.Under anesthesia, 0.2 μl of a 4% solution of FG is administered bystereotactic injection (1 mm anterior from bregma, 3 mm lateral, and 4.5mm ventral from dura into both striata). One week after FG injection,the rats receive a stereotactic injection of 6-OHDA (20 μg dissolved in4 μl saline; Sigma) into the striatum on one side of the brain, at thesame coordinates as the FG injection. Hsp70 inducing compounds of theinvention can be administered by intravenous or oral administration atdoses of 10-100 mg per kg of body weight. The compounds can be given atthe time of 6-OHDA injection or some time (2-4 weeks, for example)subsequent to 6-OHDA treatment. Rats are sacrificed 8 and 16 weeks after6-OHDA injection. The endpoints of this model are 1) behavioral changesas monitored in-life at various times by assessment of turning(rotational) behavior using classical neurological read-out, and 2) thebrain is removed after sacrifice, thin sections are made using acryostat, and immunohistochemistry is performed as described in Zigmondand Stricker (1984). Efficacy of the Hsp70 inducing compounds of theinvention is demonstrated by a decrease in rotational behavior as wellas a reduction in the loss of nigral dopaminergic neurons.

Example 7 Alzheimer's Disease (AD)

There are several transgenic mouse models of AD. One such model that iswidely used to test the efficacy of drugs in AD was described byHolcomb, et al. (Holcomb, L., Gordon, M. N., McGowan, E., Yu, X.,Benkovic, S., Jantzen, P., Wright, K., Saad, I., Mueller, R., Morgan,D., Sanders, S., Zehr, C., O'Campo, K., Hardy, J., Prada, C. M., Eckman,C., Younkin, S., Hsiao, K., and Duff, K. (1998) AcceleratedAlzheimer-type phenotype in transgenic mice carrying both mutant amyloidprecursor protein and presenilin 1 transgenes. Nature Medicine4:97-100). This model contains two different genes associated with AD.One is a mutation in the amyloid precursor protein (APP). The mutant APP(K670N, M671L) transgenic line, Tg2576, has elevated amyloidbeta-protein levels at an early age, and, later, develops extracellularAD-type A beta deposits in the brain. The other gene is a mutatedpresenilin-1 (PS1) gene. The doubly transgenic progeny from a crossbetween Tg2576 and the PS1 mutant PS1M146L transgenic line develop largenumbers of fibrillar A beta deposits in cerebral cortex and hippocampusfar earlier than their singly transgenic Tg2576 mice.

Hsp70 inducing compounds of the invention can be dosed in mice atvarious times. The age of mice at the start of drug dosing may bevaried. For example, a treatment starting time may be at 3 months ofage, a time at which the brain deposits are first detectable. The dosemay be, for example, 10 to 100 mg/kg body weight administered once perweek or three times per week by the oral or intravenous route. Theeffect of drug treatment can be assessed by measuring AD-type depositsin the brain as well as by assessing function of the mice in a mazetest.

Example 8 Measurement of Heat Shock Protein 70 (Hsp70)

Plasma Hsp70 can be measured by a sandwich ELISA kit (StressgenBioreagents Victoria, British Columbia, CANADA) according to a modifiedprotocol in house. In brief, Hsp70 in plasma specimens and serialconcentrations of Hsp70 standard are captured onto 96-well plate onwhich anti-Hsp70 antibody was coated. Then captured Hsp70 is detectedwith a biotinylated anti-Hsp70 antibody followed by incubation witheuropium-conjugated streptavidin. After each incubation unboundmaterials are removed by washing. Finally, antibody-Hsp70 complex wasmeasured by time resolved fluorometry of europium. Concentration ofHsp70 is calculated from a standard curve.

Example 9 Measurement of Natural Killer Cell Cytotoxic Activity

The following procedure can be employed to assay NK cell activity in asubject. The procedure is adapted from Kantakamalakul W, Jaroenpool J,Pattanapanyasat K. A novel enhanced green fluorescent protein(EGFP)-K562 flow cytometric method for measuring natural killer (NK)cell cytotoxic activity. J Immunol Methods. 2003 Jan. 15; 272:189-197,the entire teachings of which are incorporated herein by reference.

Materials and methods: Human erythroleukaemic cell line, K562, isobtained from American Type Culture Collection (CCL-243, American TypeCulture Collection, Manassas, Va.), and cultured in RPMI-1640 medium(Cat#11875-093Gibco Invitrogen Corp, Carlsbad, Calif.) supplemented with10% heat inactivated fetal calf serum (Gibco), 2 mM L-glutamin, 100μg/ml streptomycin and 100 IU/ml penicillin at 37° C. with 5% CO₂. K562cells are transduced with retroviral vector which encode greenfluorescent protein (eGFP). Stable cell line is selected withantibiotic, G418. About 99.6% G418 resistant cells are eGFP positiveafter section.

The subject's peripheral blood mononuclear cells (PBMCs) are prepared byclinical study sites and received in BD Vacutainer Cell Preparation Tubewith sodium heparin (Product Number: 362753, Becton Dickinson, FranklinLakes, N.J.).

Two-fold serial dilution of 800 μl effector cells (patient's PBMC)starting at concentration of 1×10⁶ cells/mL are put into four individualpolystyrene 12×75-mm tubes. Log phase growing target cells (K562/eGFP)are adjusted with growth medium (RPMI-1640) to a concentration of 1×10⁵cells/mL and 100 μL targets then added into the tubes to provideeffector/target (E/T) ratios of 80:1, 40:1, 20:1, 10:1. Effector cellsalone and target cells alone are used as controls. All tubes areincubated at 37° C. with 5% CO₂ for about 3.5 hr. Ten microliters ofpropidium iodide (PI) at a concentration of 1 mg/mL is added to eachtube including effector and target control tubes and then incubated atroom temperature for 15 min.

Cytotoxic activity is analyzed with a FACSCalibur flow cytometer (BectonDickinson). Linear amplification of the forward and side scatter(FSC/SSC) signals, as well as logarithmic amplification of eGFP and PIemission in green and red fluorescence is obtained. Ten thousand eventsper sample tube with no gating for acquisition are collected foranalysis. Data analysis for two-parameter dot plots for eGFP versus PIis performed using CELLQuest (Becton Dickinson Biosciences) software toenumerate live and dead target cells. Debris and dead cells are excludedby setting a threshold of forward light scatter.

Example 10 Inhibition of HUVEC Cell Migration

To examine if the compounds of the invention affect endothelial cellfunction, an in vitro human umbilical vein endothelial cell (HUVEC)migration assay is performed in the presence of a compound of theinvention. HUVEC cells (passage number 4) are cultured on 12-well platesand time-lapse imaging is performed with the live cell imaging system onan inverted microscope supplied with 6-7% CO₂. The temperature is keptat 37° C. Images are taken every 30 minutes using the 2× objective forup to 106 hr or every 60 seconds using the 20× objective for 30 min.Confluent HUVEC cultures are scraped similarly to make a blank area,followed by culturing in HUVEC medium for 15 hr without treatment. Themigration areas, which are imaged as time-lapse sequences for each well,are used as a basis to standardize/correct migration rates. Then,migration of cells under different treatments is imaged at the same timeto generate time-lapse image sequences for each well. Time-lapse moviesare further analyzed by measuring areas that are covered by migratingcells. During experiments, HUVEC cells are activated by the presence ofVEGF and basic FGF. Compounds of the invention (e.g. 100 nM and 1 μM)are expected to completely block migration of HUVEC cells to the blankarea, indicating that compounds of the invention possesses potentinhibitory effect on the migration of activated HUVEC cell in vitroinduced by VEGF and basic FGF.

It is also possible to track HUVEC behavior during above treatments. Itis expected that HUVEC cells will begin to shrink after 24 hr treatmentwith compounds of the invention.

Example 11 Enhanced VE-Cadherin Junctions of HUVEC Cells

An immunofluorescence study is performed by using anti-VE-cadherinantibodies to examine VE-cadherin junctions between HUVEC cells. HUVECcells are treated with DMSO or a compound of the invention (e.g. 10, 100and 1000 nM) for 24 hrs and fixed for immunostaining. DMSO concentrationis 1:100 for all treatments. To boost the immunofluorescence signal,cells are stained with a mixture of 2 polyclonal anti-human VE-cadherinAbs followed by staining with a mixture of fluorescent secondaryantibodies. It is expected that with compounds of the invention,VE-cadherin staining will be extremely strong in cell-cell junctionregions, but not the non-contacted regions compared to that in DMSOtreated cultures. Compounds of the invention are expected to enhance theassembly of cell-cell junctions of activated human endothelial cells,likely through induction of the accumulation of VE-cadherin molecules atthe junctions. This effect could result in limited motility of the cellsand reducing permeability of the endothelium, thus contributing to thecell migration inhibition and the potential anti-angiogenesis effect ofcompounds of the invention.

The relevant teachings of all publications cited herein that have notexplicitly been incorporated herein by reference, are incorporatedherein by reference in their entirety.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by formula (I):

or a pharmaceutically acceptable salt or prodrug thereof, wherein: X₁ is C(R₉)₂ or N—X₂; X₂ is R₁₇, —OR₁₇, —N(R₂₆)₂, or —NHR₂₇; Z₁, Z₂, Z₃, and Z₄ are each independently O or S; R₁ and R₂ are independently an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, halo, nitro, cyano, guanidino, —OR₁₇, —NR₁₉R₂₀, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇, —S(O)_(p)R₁₇, —OS(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or —S(O)_(p)NR₁₉R₂₀; R₃ and R₄ are independently —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl or an optionally substituted heteroaryl; R₅ and R₆ are independently —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl or an optionally substituted heteroaryl; each R₉ is independently —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, halo, nitro, cyano, guanidino, —OR₁₇, —NR₄₀R₄₁, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —OP(O)(OR₁₇)₂, —SP(O)(OR₁₇)₂, —SR₁₇, —S(O)_(p)R₁₇, —OS(O)_(p)R₁₇, —S(O)_(p)OR₁₇, —NR₁₈S(O)_(p)R₁₇, or —S(O)_(p)NR₁₉R₂₀; R₁₇ and R₁₈, for each occurrence, are, independently, —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted aralkyl, or an optionally substituted heteraralkyl; R₁₉ and R₂₀, for each occurrence, are independently —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted aralkyl, or an optionally substituted heteraralkyl; or R₁₉ and R₂₀, taken together with the nitrogen to which they are attached, form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; R₂₆ is an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted aralkyl, or an optionally substituted heteraralkyl; R₂₇ is —H, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted aralkyl, or an optionally substituted heteraralkyl, provided that R₂₇ is not an optionally substituted phenyl; R₄₀ and R₄₁, for each occurrence, are independently H, —OR₁₇, —NR₁₉R₂₀, —C(O)R₁₇, —C(O)OR₁₇, —OC(O)R₁₇, —C(O)NR₁₉R₂₀, —NR₁₈C(O)R₁₇, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl, an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted aralkyl, or an optionally substituted heteraralkyl, or R₄₀ and R₄₁, taken together with the nitrogen to which they are attached, form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; and p is 1 or 2; and provided that the compound is not propanedioic acid, (2-methylpropylidene)-, bis[2-[(phenylamino)thioxomethyl]hydrazide]. 2-5. (canceled)
 6. The compound of claim 1, wherein: R₁ and R₂ are each an optionally substituted aryl or an optionally substituted heteroaryl; and R₃ and R₄ are each an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 7. The compound of claim 6, wherein R₅ is —H and R₆ is —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; and R₃ and R₄ are each an alkyl group.
 8. The compound of claim 7, wherein R₁ and R₂ are each a substituted or unsubstituted phenyl group and R₃ and R₄ are each methyl or ethyl.
 9. (canceled)
 10. The compound of claim 1, wherein R₁ and R₂ are both an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; R₅ is —H; and R₆ is —H or an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 11. The compound of claim 10, wherein R₁ and R₂ are both a C3-C8 cycloalkyl group optionally substituted with at least one alkyl group; R₃ and R₄ are both an alkyl group; and R₆ is —H or methyl.
 12. The compound of claim 11, wherein R₁ and R₂ are both cyclopropyl or 1-methylcyclopropyl. 13-15. (canceled)
 16. The compound of claim 1, wherein the compound is represented by formula (III):

17-20. (canceled)
 21. The compound of claim 16, wherein: R₁ and R₂ are each an optionally substituted aryl or an optionally substituted heteroaryl; and R₃ and R₄ are each an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 22. The compound of claim 21, wherein R₅ is —H and R₆ is —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; and R₃ and R₄ are each an alkyl group.
 23. The compound of claim 22, wherein R₁ and R₂ are each a substituted or unsubstituted phenyl group and R₃ and R₄ are each methyl or ethyl. 24-25. (canceled)
 26. The compound of claim 16, wherein R₁ and R₂ are both an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; R₅ is —H; and R₆ is —H or an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 27. The compound of claim 26, wherein R₁ and R₂ are both a C3-C8 cycloalkyl group optionally substituted with at least one alkyl group; R₃ and R₄ are both an alkyl group; and R₆ is —H or methyl.
 28. The compound of claim 27, wherein R₁ and R₂ are both cyclopropyl or 1-methylcyclopropyl. 29-30. (canceled)
 31. The compound of claim 1, wherein the compound is represented by formula (V):

32-35. (canceled)
 36. The compound of claim 31, wherein: R₁ and R₂ are each an optionally substituted aryl or an optionally substituted heteroaryl; and R₃ and R₄ are each an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 37. The compound of claim 36, wherein R₅ is —H and R₆ is —H, an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; and R₃ and R₄ are each an alkyl group.
 38. The compound of claim 37, wherein R₁ and R₂ are each a substituted or unsubstituted phenyl group and R₃ and R₄ are each methyl or ethyl. 39-40. (canceled)
 41. The compound of claim 31, wherein R₁ and R₂ are both an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl; R₅ is —H; and R₆ is —H or an optionally substituted alkyl, an optionally substituted alkenyl, an optionally substituted alkynyl, an optionally substituted cycloalkyl, an optionally substituted cycloalkenyl, an optionally substituted heterocyclyl.
 42. The compound of claim 41, wherein R₁ and R₂ are both a C3-C8 cycloalkyl group optionally substituted with at least one alkyl group; R₃ and R₄ are both an alkyl group; and R₆ is —H or methyl.
 43. The compound of claim 42, wherein R₁ and R₂ are both cyclopropyl or 1-methylcyclopropyl. 44-45. (canceled)
 46. The compound of claim 1, wherein the compound is represented by formula (VII):


47. The compound of claim 46, wherein X₁ is C(R₉)₂.
 48. The compound of claim 47, wherein X₁ is N—X₂.
 49. A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a compound of claim
 1. 50-52. (canceled)
 53. A method of treating a subject with a cancer, said method comprising administering to the subject an effective amount of a compound of claim
 1. 54-63. (canceled) 